Progress in Cell Cycle Research, Vol.

3, 77-97 , (1997)
(Meijer, L., Guidet, S. and Philippe, M., eds.) Plenum Press, New York, USA

chapter 7
Protein kinase CK2 ("casein kinase-2") and
its implication in cell division and proliferation
Lorenzo A. Pinna' and Flavio Meggio

Dipartimento di Chimica Biologica and Centro di Studio delle Biomembrane

del Consiglio Nazionale delle Ricerche, Universitii di Padova, via Trieste 75, 35121 Padova, Italy.
"To whom correspondence should be addressed

Protein kinase CK2 (also termed casein kinase-2 or -IT) is a ubiquitous Ser/Thr-specific protein kinase
required for viability and for cell cycle progression. CK2 is especially elevated in proliferating tissues,
either normal or transformed, and the expression of its catalytic subunit in transgenic mice is causative of
lymphomas. CK2 is highly pleiotropic: more than 160 proteins phosphorylated by it at sites specified by
multiple acidic residues are known. Despite its heterotetrameric structure generally composed by two
catalytic (a and/or a') and two non catalytic ~-subunits, the regulation of CK2 is still enigmatic. A number of
functional features of the ~-subunit which could cooperate to the modulation of CK2 targeting/activity will
be discussed.

INTRODUCTION apparatus casein kinase (re-named G-CK) being that

The acronym "CK2" derives from "casein kinase-
2", a term coined after the ability of this enzyme to
phosphorylate casein readily in vitro. Although
widely used, this term is somewhat misleading as
casein most likely does not belong to the endless list
of substrates of this pleiotropic protein kinase. Hence
the proposal to de-emphasise the concept of "casein
phosphorylation" by using the abbreviation "CK2"
instead of the full description "casein kinase-2" [1].
Although casein can be more or less readily
phosphorylated in vitro by a variety of protein
kinases, including some tyroSine kinases, three
classes of Ser /Thr specific protein kinases display a
marked preference for casein over the majority of
other protein substrates and were therefore named
"casein kinases". Besides CK2 these are protein
kinase CKI ("casein kinase-I"), another ubiquitous
and pleiotropic enzyme structurally and
physiologically unrelated to casein [2], and bona fide
casein kinase specifically expressed in lactating
mammary gland where it is committed to the
phosphorylation of newly synthesised casein
molecules (see table 1). Until recently this latter
enzyme was believed to represent a highly dedicated
kinase specifically implicated in the processing of
casein during its secretion in the Golgi apparatus of
the mammary gland (hence its acronym "GEF-CK",
Golgi enriched fraction casein kinase [3]). It has
recently been shown however that a protein kinase
biochemically indistinguishable from mammary
gland GEF-CK is also present in the Golgi apparatus
of rat liver, spleen and, to a lesser extent, kidney and
brain [4]. It has to be concluded therefore that all the
three classes of "casein kinases" are ubiquitous and
pleiotropic enzymes, a unique feature of the Golgi
of counting casein among its natural substrates,
together with others which are currently only
conjectural [4]. In contrast to G-CK, which is
localised in the Golgi apparatus and absent from the
rest of the cell, CK2 and CKI are hardly detectable in
the Golgi apparatus [4]. CK2 is present in several
subcellular fractions, especially the nucleus, where it
reaches its highest concentration [5-7]. Additional
features that differentiate CK2 from the other two
classes of "casein kinase" are its ability to use GTP,
besides ATP, as phosphate donor, its
heterotetrameric structure, rare among protein
kinases and its predilection for phospho acceptor
residues having clusters of acidic amino acids on
their C-terminal side. Such a remarkable site
specificity is dictated by a minimum consensus
sequence, S/T-X-X-E/D/Sp/Yp which is similar to,
yet definitely distinct from, the one recognised by G-
CK, S/T-X-E/Sp (see table 1).
The ubiquity of CK2, in conjunction with its
requirement for viability [8], unmatched pleiotropy
[9] and recurrent observations linking CK2 over-
expression with neoplastic growth [10, 11] have
attracted increasing attention to CK2, which in the
last decade has been dealt with in several review
articles [1, 2, 9, 10, 12-14]. Despite these efforts and
the large amount of information available, CK2
remains a somewhat enigmatic kinase, whose crucial
role in cell regulation and whose implication in
signalling, gene expression and proliferation is based
more on a wide variety of coincidental observations
rather than on clear-cut evidence of cause-and-effect.
It is particularly striking that the mode of regulation
of CK2 remains obscure, and that CK2 does not take
part in any of the networks of signal transduction

77
1. A. PINNA AND F. MEGGIO
Table 1. DIstinctive properties of the three classes of "casein kinases".
Tissue distribution Ubiquitous
Main subcellular localization Golgi
Natural targets Casein and others
PKgroup ? CKl/HHR25
(Hanks and Hunter)
Mr holoenzyme (kDa) 400-500
Subunit composition ? Monomeric
Phosphate donor ATP
Consensus sequence SxE/Sp
pathways which have been elucidated in recent
years. Consequently the inclusion of CK2 in the
category of 'growth related protein kinases' [15]
should continue to be regarded with healthy
scepticism until such time as it has a more rigorous
experimental basis.
Our present aim is to scrutinise the molecular
features and functional properties of CK2 that could
help to understand why this protein kinase, essential
for viability and causative of tumors, has been
committed throughout evolution to phosphorylating
such a wide variety of cellular components. In
particular we shall focus on structural and functional
features that may reflect or underlie the mode of
regulation of CK2, and compare CK2 with other
'growth related' kinases, particularly with the cyclin-
dependent kinases (CDKs) which are closely related
to CK2 within the phylogenetic tree of protein
kinases, and share with CK2 the use (rare among
protein kinases) of 'regulatory subunits', but
nevertheless exhibit quite distinct, and in some
respects opposite, biochemical properties.
STRUCTURAL FEATURES
Subunit composition
In the vast majority of cases CK2 isolated from a
wide variety of organisms and tissues has been
described as a spontaneously active heterotetramer
with a Mr of approximately 130,000, composed of
two catalytic subunits (a and/or a' of Mr 42-44 and
38 kDa, respectively) and two non catalytic p-
subunits whose Mr in animals is approximately 26
kDa). The catalytic subunits, which are the product
of distinct genes, share high homology, a' being
shorter than a at its C-terminus. In the human
genome there are two loci encoding the a-subunit,
and a single locus for each of the a' and p-subunits.
The structures of the human a and p, S. cerevisiae a
and a', C. elegans a and p, mouse a and Theileria paroa
a genes are known (reviewed and/or referenced in
[14]). The actual occurrence of all the three predicted
78
CKl CK2
Ubiquitous Ubiquitous
Microsomes Nuclei
Cytosol Cytosol
Many Many
(not casein) (not casein)
CMGC
40-60 130
a2P2/ aa'P2/ a'2P2
ATP ATP/GTP
SpxxSLIB SLIxxE/D /YpSp
tetramers, a2P2, a'2P2 and a dP2 has been
documented [7, 16]. In S. cerevisiae two kinds of P
subunits exist, P and po, both larger than animal P
subunits and expressed by separate genes [17]. A
similar situation applies to the plant Arabidopsis
thaliana [18]. In S. cerevisiae it seems likely that CK2
holoenzyme is an obligatory heterotetramer of all
four subunits [18a]. In contrast monomeric CK2
enzymes composed of just a single catalytic subunit
have been isolated from Zea mays [19] and
Dictyostelium [20], where no protein equivalent of P
has yet been found. Pertinent to this may be also the
vexed question of whether isolated catalytic subunits
might exist under certain circumstances. Failure to
isolate monomeric isoforms of CK2 from tissues
where the heterotetramers are present and co-
localisation of catalytic and p-subunits detected by
immunofluorescence microscopy in embryo
fibroblasts, virally transformed chicken hepatoma
cells and HeLa cells [6] would argue against the
presence of catalytic subunit not combined with the
p.-subunits. However there are also observations
supporting this possibility, notably the finding that
the majority of the a-subunit in the nuclei of
Chironomus ten tans [21] is complexed to internuclear
components devoid of its p.-subunit counterpart and
the intriguing finding that over-expression of p in
COS-l cells evoked an increase of CK2 catalytic
activity suggestive of the recruitment of catalytic
subunits, from a storage pool where they are poorly
active, to give more active heterotetramers [22]. The
functional implications of this and other observations
will be discussed below (see section entitled
"Regulation").
Structure of catalytic (ala') subunits
The primary structure of CK2 catalytic subunits
(either a or a') displays the features that have
become the hallmarks of the large superfamily of
protein kinases. In particular 15 highly conserved
amino acids with three or fewer variant residues
(identified from the analysis of 75 Ser /Thr protein
kinases [23]) are all present in CK2 catalytic subunits
 CHAPTER 7I PROTEIN KINASE CK2

(underlined in fig. IA) Most of these residues are
known to cluster around the active site and are
involved in ATP binding and phosphotransfer [24],
which are properties common to all protein kinases,
whether Ser /Thr or Tyr specific. These highly
conserved residues allow one to recognize within the
CK2 (l I (l' structures the canonical twelve
subdomains that define most of the members of the
family. A more detailed sequence analysis shows
that CK2 catalytic subunits share the highest
similarity with cyclin dependent protein kinases
(CDKs), mitogen-activated protein kinases (MAP
kinases) and Glycogen Synthase Kinase-3, together
forming one of the main branches of the
phylogenetic tree, the so called CMCG group [25]. A
common feature, shared by all members of this
group is the presence of two inserts, a 'small' one
between subdomains IX and X and a 'large' one
between subdomain X and XI (see fig. IA). It should
be noted, however, that the percentage identity of the
CK2 catalytic subunits with their closest relatives
does not exceed 33%, and that many of the peculiar
features conserved across the (ll (l' subunits from
different species (highlighted in fig. 1) are not
present in the other members of the CMCG group;
notably these share with the majority of protein
kinases two residues (A70 and FI85 in PKA) which
are replaced in CK2 catalytic subunits by V and W,
respectively; and the characteristic KKKKIKR basic
stretch preceding in CK2 the conserved Glu that
defines subdomain III is not basic in CDKs
whichever alignment is followed (fig. IC). By the
canonical multiple alignment used by Hanks and
Quinn [23] this basic segment matches the PSTAIR
motif of CDKs implicated in cyclin binding. If a
manual alignment is followed based on the greater
similarity between CK2 and PKA and CDKs, the
basic block of CK2 aligns with an acidic sequence of
CDK2 (TETEG) which could account for the selection
of a basic, instead of acidic, determinant at position

A
I B (Glyloop)
MSGPVPSRARVYTDVNTHRPREYWDYESHVVEWGNQDDYQL~ 50 PKA KTL"TGSFGR
glycine MAPK SYI"E"AYGM
CDK2 AKI"E,,'l'YGV
II III IV CK2 RKL"R"ItYSES2
...
loop
.......... .
S~INI~ILKPVJaaOuKRIIKJ:LENLRGGPNJ:ITLADI

.\&betrate biAdlDg, IlL'

100

J:8I./BSPJO/heparin billdillg'
interaction with P dowa-regulatory 4c.aill C (Subd. IlIIlI connection)
V VIa PKA KILDKQKVVKLKQIEHT~
VKDPVSRTPALVFEHVNNTDFKQLYQTLTDYDIRFYMYEILKALDYCHSM 150 MAPK KKISPFEHQ--TYCQRTLR~
CDK2 KKIRLDTETE-GVPSTAIR~
CK2 KILKPVKKKKIK------~81
Vlb VII
G~qvMrDBEHRXLRLI~FYHPGQEYNVRVASRYFKGl! 200
catalytic loop activatioza
loop
loop pel
D (catalytic loop)

VIII IX X PKA Y~LKPEULL

ILLVDYQMYDYS~S~MIFRKEPFFHGHmfYDQLVRIAKVLG MAPK H~LKPSNLL
250
CDK2 H~LKPQNLL
CK2 HR~VliPHNVM163

..........................
TEDLYDYIDKYNIELDPRFNDILGRHSRKRWERFVHSENQRLVSPBALDF
aajor in•• rt
300
E (activation loop plus loop p+1)
XI PKA GRT------WTLCGTPEYLA~
LDKLLRYDHQSBLTAREAMEHPYFYTVVKDQARMGSSSMPGGSTPVSSAN 350 MAPK PDHDHTGFLTEYVATRWYRA~
CDK2 VPVRTY - - -THEVVTLWYRA~
CK2 HPGQEY---NVRVASRYFKG~201

~mSGISSVPTPSPLGPLAGSPVlAAANPLGM?VPAAAGAQQ 391

Figure 1. Amino acid sequence and main structural features of human CK2 a-subunit.
In A the primary structure is reported. Bold typing denotes residues that are conserved in at least 17 out of 19 sequences of a and a'
reported in the EMBL data bank from H. sapiens (M55265, M55268), B. taurus (M93665), C. gallus (M59456, M59457), O. cuniculus (M98451),
R. norvegicus (L15618), D. rerio (X99964), X. Ioeuis (X62375), D. melanogaster (M16534), c. elegans 005274), D. discoideum (L05535), S. cerevisiae
(M22473, M33759), S. pombe (X74275), T. porva (M92084), A. tlulliana (010246, 010247), Z. mays (X61387). Highly conserved amino acids
present in at least 72 out of 75 Ser /Thr protein kinases [23] are underlined. These define subdomains common to all protein kinases
(indicated by Roman numbers). Series of bullets below the sequence define knoym functional regions, as indicated.
In B, C, 0 and E the glycine loop, the basic stretch connecting subdomain II and III, the catalytic loop and the activation plus p+ 1 loops,
respectively, are compared with the homologous sequences of PKA, MAPK and CDK2 (alignment as in [206]). Invariant residues are
underlined. Bold typing denotes residues whose function is discussed in the text (see sections entitled, "Structure of catalytic (a/ a')
subunits", "Catalytic properties and specificity" and "Regulation").

79
L. A. PINNA AND F. MEGGIO
n+3 by CDKs as compared to CK2 [26]. As discussed
below, the CK2 K74-77 basic quartet is implicated in
both substrate recognition (position n+3) and
interaction with the ~suburiit. Thus, either the
PSTAIR or the TETEG alignments would be
plausible, assuming that homologous residues play
similar functions.
In fig. lA bold type denotes those residues that
are conserved in at least 17 out of the 19 sequences of
a and a' considered; Roman numbers refer to the
subdomains common to all protein kinases.
Additional structural features are also indicated. The
following regions deserve comment.
The Gly loop (also termed phosphate anchor, as in all
kinases it makes contact with the ~ phosphate of
bound ATP). The first two glycines out of the three of
the motif G-X-G-X-X-G are conserved, the third one
being replaced by a hydrophilic residue, serine, a
feature that might confer ATPase activity by bringing
water molecules in contact with ATP [27]. Two
features of this loop are notable: a lysyl residue (K49)
which has been shown by mutational analysis to
contribute to the recognition of the acidic
determinant at pOSition n+2 in peptide substrates
(Sarno et al., unpublished); and a tyrosine (Y50)
which is homologous to the regulatory residue Y15
of CDK, the phosphorylation of which (as well as
that of preceding T14) suppresses the catalytic
activity of cyclin-dependent kinases. In contrast, this
tyrosine is replaced by a non-phosphorylatable
residue (usually phenylalanine) in PKA and in most
kinases other than CDKs and CK2 (see fig. IB). It is
curious that in CK2 from Theileria parua, the agent of
a leukemia like disease in cattle, Y50 is replaced by a
phenylalanine [28].
The 74-80 basic stretch. This sequence, which is
probably the most striking hallmark of CK2 (see fig.
IC), is situated just upstream from the conserved
glutamic acid that defines subdomain III and just
downstream from a series of gaps with respect to the
canonical multiple alignment [23]. However,
alternative alignments are also possible [26, 29] based
on greater homology with PKA and CDKs.
Whichever alignment is adopted, such a high
concentration of adjacent basic residues is
unmatched among protein kinases in this region.
Mutational analyses have shown that the first part of
this cluster (K74 to K77) [30-32], but not the
subsequent K79 , R80 and K83 [32], are implicated in
CK2 inhibition by heparin. However, both the 74-77
and the 79-83 segments are implicated in substrate
recognition by interacting with the crucial
determinant at position n+3 [26]. Although it appears
that these basic residues are to some extent
interchangeable as far as substrate recognition is
concerned, K77 seemingly plays a prominent role
since its individual replacement by alanine causes a
80
dramatic increase of Km towards casein (Sarno et al.,
unpublished), while the double mutations of
K74/K75 [3D] and of K75/K76 [31] proved nearly
ineffective on the Km for casein. By analogy with the
corresponding PSTAIR sequence of CDKs (if the
canonical multiple alignment is. adopted) which
interacts with cyclins [33], the 74-80 basic sequence of
CK2 interacts with the ~subunit (Sarno et al.,
submitted). However it mediates down-regulation by
~, not the 'cyclin-like' positive effect on catalytic
activity; and this latter is actually more pronounced
with CK2a mutated in the 74-80 basic segment
(Sarno et al., submitted). Competition experiments
with heparin would suggest that the 74-77 basic
quartet also mediates interactions with HSP90 [34]
and with nucleolin [35]. Another putative function of
this segment would be to target CK2 to the nucleus,
as it contains a canonical nuclear localisation signal
[36].
The catalytic loop and the PP2A binding motif. This
short segment spanning the highly conserved Asp
and Asn residues that define subdomain VIB (see fig
lA) in CK2 is almost identical to those in PKA and
CDKs (see fig. ID), with the important exception,
however, of a basic residue (HI60) replacing the
acidic one (E170 in PKA) found in the majority of
protein kinases. PKA El70 interacts with the crucial
arginine present at position -2 in most PKA
substrates. The same role is played by acidic residues
homologous to E170 in other basophilic kinases [37].
Its replacement by a histidine in CK2 appears
indicative of the acidophilic nature of this kinase,
although position n-2 is not so important in CK2
substrates. Mutational studies have corroborated this
interpretation [38]. Just downstream from the
catalytic loop is a sequence motif conserved in
vertebrate and Drosophila CK2a (H-F-H/N-R-K-L),
although not in CK2a from other species. This motif
is also found in the region of SV40 required for
binding protein phosphatase 2A, and has been
shown to account for the ability of CK2a to bind to
PP2A in vitro [39].
The subdomain VII-VIII connection (activation loop
and loop p+1). The region between the 'DFG' and
'APE' triplets that define subdomains VII and VIII in
most protein kinases (both altered in CK2, where
they are DWG and ~PE, respectively) contains two
important functional elements termed the 'activation
loop' (or 'T-Ioop' after the presence of a threonyl
residue that is constitutively phosphorylated in
active PKA) and 'loop p+l'. The activation loop
includes residue(s) the phosphorylation of which
(either autocatalytic or promoted by other kinases)
correlates with increase of activity in most, albeit not
all, protein kinases [40]. In particular in CDK2, for
which the crystal structures of both the inactive
catalytic subunit [41] and the partially active
complex with cyclin A [33] are available, it has been

shown that the activation loop blocks the entrance to
the catalytic cleft in the free kinase, while in the
complex with cyclin A it adopts a different
conformation in which its constituent residue Thr-
160 is exposed and available for phosphorylation.
This latter leads to full activation of the kinase. As
outlined in Fig. IE, CK2ex, in contrast to CDKs and
MAP kinases (belonging to the same CMGC group),
does not contain either threonine or serine in its
activation loop nor has phosphorylation in this
region ever been reported (see section entitled
"Regulation"). These observations, in conjunction
with the finding that free CK2 catalytic subunit (in
contrast to the free catalytic subunit of CDKs) is
spontaneously active, suggest that the activation
loop of CK2 is displaced from the catalytic cleft and
does not hamper enzymatic activity.
The 'p+ 1 loop', which is adjacent to the C-
terminal edge of the activation loop, derives its name
from the fact that in PKA it includes three
hydrophobic residues (L198, P202 and L205) which
form a pocket surrounding a leucine at position p+l
in the inhibitor peptide co-crystallized with the
catalytic subunit [42]. These hydrophobic residues
are conserved or conservatively replaced in most
Ser /Thr protein kinases. In contrast they are replaced
by basic residues (RI91, R195 and K198) in CK2
which strongly selects acidic residues at position n+1
(see fig. IE). Their simultaneous replacement with
alanines gives rise to a mutant which is severely
defective in substrate recognition, being unable to
bind the acidic determinant at n+ 1 [26]. An almost
comparable effect is observed if K198 alone is
mutated (Sarno et al., unpublished). It is interesting
that in all the other members of the CMGC group,
which select a proline at position n+ 1 [37], the
residue homologous to K198 is an arginine, which
has been shown by space-filling modelling to match
well a prolyl residue [43].
The 'small' and 'large' insert regions. These two
inserts preceding and following subdomain X, are a
hall-mark of protein kinases of the CMGC group.
The large insert, despite its extension, does not alter
the overall architecture of either CDK2 [41] or ERK-2
[44] as compared to the prototype kinase, PKA,
which lacks this insert. Rather it gives rise to a
protuberance in the lower part of the large lobe,
whose overall structure otherwise overlaps the large
lobe of PKA. This structural element does not make
important contacts with cyclin A and accordingly
deletion of the large insert from cdc2 did not abolish
activity or cyclin binding [45]. Likewise mutations
affecting the same insert in CK2 [46, 26] do not alter
catalytic efficiency nor the ability to bind the p-
subunit. However it is interesting that deletion of the
large insert from cdc2 prevents the binding of suc1,
another component of the cdc2 active complex [47],
suggesting that it also may be used by CK2 for
81
CHAPTER 7/ PROTEIN KINASE CK2
interacting with ligands, as yet unknown.
Alternatively it has been suggested that this large
insertion in MAP kinases might provide an extended
region of interaction with protein substrates which
bind better than short peptides to these kinases [44].
However this seems unlikely to be the case for CK2,
since it generally phosphorylates peptide substrates
as efficiently as the protein from which they are
derived [48]. It is possible, however, that the large
insert might help CK2 to recruit particular substrates
lacking a strong consensus sequence at their
phosphoacceptor sites.
In contrast to the large insert, the small one,
which in CK2 includes two histidinyl residues, may
indeed be implicated in peptide substrate recognition
since its mutation to that present in CDK2 caused a
significant increase in the Km value for the peptide
substrate R3DJSDJ [46].
The CDK phosphorylation sites in the C-terminal tail
of the ex-subunit. The C-terminal segment of the
vertebrate ex-subunit, which lies outside the catalytic
core, is poorly conserved in the ex-subunits of lower
organisms, and is absent from the ex'-subunits. This
tail contains a number of Ser /Thr residues which
display the consensus for and are phosphorylated by
cyclin-dependent kinases both in vitro and in vivo
[49]. Although such a phosphorylation does not
correlate with any overt change of activity, it is
possible that it may modulate other properties of the
ex-subunit, e.g. assembly with the p-subunit.
Pertinent to this may be the observations that ex'-
subunits, lacking the phospho acceptor sites,
assemble into holoenzyme more rapidly than ex-
subunits in vivo [7], and are more susceptible than ex-
subunits to up-regulation by the p-subunit [SO).
Structure of the p-subunit
The p-subunit of protein kinase CK2 does not
share similarity with the catalytic subunits of protein
kinases or with any other protein known, with the
notable exception of the product of the 'Stellate' gene
of Drosophila, which exhibits 38% identity to the p-
subunit from the same insect. Typically p-subunits
from vertebrates and insects are composed of 215
amino acids exhibiting high sequence conservation
(88% identity between human and Drosophila). The P-
subunit from the nematode C. elegans still displays
80% identity to human p in amino acids 1-200, but
has a unique 19 amino acid C-terminal extension
beyond residue 215. The p-subunits present in yeasts
and plants, besides being more divergent also tend to
display larger Mr owing to N-terminal extensions
and/or to inserts [51].
The primary structure of the human p-subunit is
shown in fig. 2, with residues conserved in all known
animal p-subunits shown in bold type. Underlining
denotes those residues that are also conserved or
L. A. PINNA AND F. MEGGIO
IISSSBZVStlI~<mKDIL~HIRQAL 50
••••• •••
autopboapborylatioa
bill4iDSJ of IIoppUO

... .......... .
~PDEET.BJlRPNQSDLlIQAABMLYGLlBARnLi'NRa...~ 100

• • •
ZiDC fiDSJ.~ ziDC fiDIJ.~

••••••••••••• .........................
phyaical a-~ at~ctu~al iDt.~actioD
A-Ref bill4ing

LQAASNFKSPVKTIR 215
•
cdk pboapborylatioD

Figure 2. Amino add sequence of human CK2IJ-slibuniL

Bold typing denote residues conserved in 6 vertebrate (ff. sapiens, XI6312; G. gallus, M59458; O. cuniculus, M98450; R. norvegicus, L15619; S.
scro{tl, X56503; X. /oeois, X62376) and in Drosophila (MI6535) CK2 IJ-subunits found in EMBL data bank. Residues conserved or
conservatively replaced also in IJ and W-subunits from S. ceret1isiRe (U21283, U08849), S. pombe(X74274) and A. thaliana (L22563, U03984) are
underlined. Residues / sequences supposed to be implicated in specific functions or defining structural motifs (as indicated) are denoted by
bullets.

conservatively replaced in non-animal sequences. An
obvious feature of the p-subunit is the uneven
distribution of basic residues, concentrated in the C-
terminal region, and acidic residues which largely
predominate in the N-terminal region. A number of
structural elements responsible for the biochemical
properties and putative functions of the p..-subunit
have been mapped. It would appear that the N-
terminal region contains elements that are mainly
responsible for intrinsic down-regulation of CK2 and
sensitivity to polycationic effectors. In particular the
elements implicated in negative regulation are a
conserved acidic stretch encompassing residues 55 to
64 [52, 53] and the autophosphorylation site at the
extreme N-terminus [54, 55]. The 55-64 segment is
also responsible for sensitivity to polylysine since
mutation of acidic residues therein attenuates the
stimulation by polylysine but not that by polyamines
[52, 53, unpublished data]. The effect of these latter
polycations appears to be mediated by a region
further from the N-terminus, extending to pro-nO
[56], where a prominent role in binding spermine is
played by Thr-72 and His-I08 [57], with contribution
from Glu-73 and Glu-77 [56]. The N-terminal 20
amino acids are also involved in the binding of
Noppl40 [58], a protein that has been shown to
shuttle between the cytoplasm and the nucleus.
The structural features that are responsible for p-
p dimerization, association with the a-subunit,
protection against denaturation and proteolysis and
up-regulation of activity are located in the C-terminal
region [52, 59]. These two distinct functional
domains of the p-subunit have been physically
dissected by generating large synthetic fragments
encompassing either the N-terminal (1-77) or C-
terminal region (155-215) from which it has been
demonstrated that the former displays only the
negative effects of the p-subunit, as judged from its
ability to abolish calmodulin phosphorylation, while
the latter protects and stimulates catalytic activity
[60]. The additional finding that peptide P[155-215]
but not its shorter derivative P[171-215] (nor the N-
terminal peptide P[I-77]) tends to dimerize and to
interact with the a-subunit, as judged from far-
western blot analysis [60] supports the view that the
155-171 segment is crucial for stabilising intersubunit
association. However, plasmon resonance methods
(BIAcore), which one would expect to be more
sensitive than far western blotting, also shows that
PU-77] interacts with a, albeit much more weakly
than full-length p and PU55-215] [Sarno et al.,
unpublished]. This is in agreement with the presence
in the fragment of the acidic cluster that binds the
basic 74-79 stretch of the a-subunit [61, Sarno et al.,
manuscript submitted]. The C-terminal domain is
also essential for the interaction with A-Raf, one of
the many cellular 'partners' of CK2: data obtained
using p mutants and the two-hybrid system
approach would indicate that both the 194-200
segment and R175 are indispensable for A-Raf
binding [62]. The extreme C-terminal region of p
contains a residue (Ser-209) which is phosphorylated
by CDKs both in vitro and in vivo [63]. This serine is

82
 CHAPTER 7/ PROTEIN KINASE CK2

A
[@

~
*
+ 20t
-+
~-+
I PBP I
B

~[155-215l

W
X -+
+ 20t
-+
&
Figure 3. Speculative representation of assemblance of CK2 subunits to give the heterotetrameric holoenzyme (A).
Hatching denotes the C-terminal region of ~-subunit reproduced in the synthetic peptide ~[155-2151 (B). The dotted moiety of the (l-
subunits indicates its active site where the N-terminal region of ~subunit Oacking in ~[155-215» exerts its negative regulation (indicated by
arrows pointing down). This can be overcome by polybasic peptides (PBP) that compete against the (l 74-80 and 191-198 basic stretches for
binding to the N-terminal acidic region of ~.

absent from the ~subunits of Drosophila, yeast and
Arabidopsis. Furthermore, human ~ can be deprived
of a (-terminal segment which includes this site
without any apparent change in its functional
properties [64].
Sequence analysis of ~ has also disclosed two
intriguing structural motifs. One is located in the N-
terminal region, just upstream from the acidic
cluster and, as pointed out by Allende and Allende
[1], is closely reminiscent of the cyclin 'destruction
box' responsible for the ubiquitin-mediated
proteolysis occurring at specific times in the cell
cycle. In the region spanning the amino and
carboxy-terminal domains there are four cysteines
009, 114, 137 and 140) whose spacing fulfils the
requirements of the potential metal binding motif
(CPX3CX22CPXC) reminiscent of the zinc finger
present in certain DNA-binding proteins [65]. It is
interesting that this motif is conserved not only in
atypical ~subunits from S. cerevisiae, and S. pombe
but even in the most distant relative of the family
[51], stellate protein, in which other features of the
~subunit, notably the auto phosphorylation site and
the down-regulatory acidic cluster, are lost. This
strongly suggests that the presence of this motif
indeed is required for function(s), still unknown,
that are shared by all the members of the
CI<2~ / stellate family. It is also conceivable that the
middle region of the ~-subunit may be implicated
in interactions with some of the many cellular
partners of this protein, which are currently being
elucidated (see section entitled "Regulation").
Quaternary structure of CK2 holoenzyme
Until the structure of CK2 catalytic subunits co-
crystallized with ~-subunits (or ~ fragments) is
solved the detailed picture of heterotetrameric CI<2
will remain a matter for conjecture. Substantial clues
about the overall architecture of the holoenzyme
have, however, been provided by independent
studies performed in three laboratories using the
two-hybrid system. These have shown that, while ~­
subunits tend to interact with each other and with
the catalytic subunits, the catalytic subunits do not
interact with one another [62, 66, 67]. Any plausible
model of the heterotetramer should therefore be
based on ~~ and ~a contacts, while a-a contacts, if
any, should be marginal. The additional finding that
the dimeric form of the (-terminal ~ fragment, ~[155-
215], mainly responsible for ~a interaction, binds
more efficiently than its monomeric form [60] would
suggest that ~ dimerization is a prerequisite for tight
binding of a. This could mean that either each
catalytic subunit needs to make contacts with both ~
subunits for high affinity association, or that
dimerization of ~ promotes conformational changes
that facilitate subsequent binding of the catalytic
subunits. These two possibilities are not mutually
exclusive. Based on these premises, and bearing in
mind that the ~-subunit is much less accessible to
proteolytic enzymes after incorporation into the
holoenzyme [68], the assembly of CK2 subunits can
be depicted by the model shown in fig. 3, in which ~-

83
L. A. PINNA AND F. MEGGIO
pand p-a but not a-a interactions are depicted, and
in which Pis placed in a 'core' position, not easily
accessible to proteolytic enzymes. It should also be
recalled that the stabilisation effect is reciprocal since
both p and a change their conformation upon
assembly, as shown by variations in CD spectra [69,
70], and reduced susceptibility to denaturation [71].
The proposed model also accounts, though from a
merely pictorial stand-point, for the finding that the
main elements responsible for either p-p or p-a
interactions are concentrated in the same region (the
C-terminal one, hatched in fig. 3). This kind of tight
interaction which also confers a higher activity and
greater stability upon a is mimicked by the C-
terminal peptide p [155-215] [60], as shown in fig. 3B.
The counterpart for this stable interaction in a/a' is
unknown. On the other hand weaker and reversible
interactions (denoted in fig. 3 by dotted lines) also
occur between acidic segments in the N-terminal
domain of P (not implicated in dimerization) and a.
These are responsible for down-regulation (denoted
by downwards arrows in fig. 3) and involve the basic
stretch of a between subdomains IT and III and the
basic residues of the p+ 1 loop (see above and fig. 1).
These residues of a are also implicated in substrate
binding, although their transient electrostatic
interaction with the N-terminal region of Pcannot
account for the tight a-p association, which does not
appear to be reversible under physiological
conditions.
FUNCI10NAL ASPECTS
Catalytic properties and specificity
The crystal structure of the catalytic subunits of
CK2 is not yet available. Nevertheless, the fact that
all of the residues that are important for catalysis in
protein kinases are conserved in CK2, and that
kinetic studies [Meggio, unpublished] are consistent
with a sequential ordered mechanism comparable to
the distantly related PKA [72] and CSK [73], support
the view that the structural organisation of the
active site and the mechanism of catalysis are not
different from those of the other members of the
kinase family. The Km (5-20 IJ.M) and Kcat (about
50 min-I) values with the best peptide substrates of
CK2 also compare fairly well with those of other
Ser /Thr protein kinases. There are however some
distinctive features of CK2 that warrant a short
comment. First, CK2 is one of the few protein kinases
which are able to use GTP, besides ATP, as
phosphate donor. Whether this aptitude has any
physiological relevance is unknown. The structural
basis for this unusual nucleotide specificity has
been partially, although not entirely, elucidated by
experiments in which the two residues that in
other kinases are almost invariably Ala and Phe
have been replaced in CK2 by Val (V66) and
tryptophan (W176), respectively. Mutation of V66
and W176 to Ala and Phe, respectively, increases
84
the GTP to ATP Km ratio [70] but does not convert
CK2 into a 'regular', strictly ATP-dependent kinase.
Second, and possibly related to its unusual
nucleotide specificity, CK2 is refractory to the very
powerful competitive inhibitor, staurosporine,
which affects the majority of other protein kinases
[74]; but is more susceptible than most other
kinases to halogenated derivatives of benzi-
midazole and benzotriazole [75-77]. These obser-
vations suggest that the ATP-binding pocket and/or
its annexes are uniquely tailored in CK2. From a
practical standpoint it may also be worth noting
that the members of a new class of selective
inhibitors of cell cycle kinases, notably 010-
moucine [78] and roscovitine [79], are ineffective
against CK2. Third, in contrast to most Ser /Thr
protein kinases, the specificity of which is
dictated by basic and hydrophobic residues and
by prolines [37], CK2 belongs to a tiny group of
Ser/Thr kinases that use acidic and/or
phosphorylated residues as specificity determinants.
The minimum consensus for CK2 is S/T-X-X-
E/D/Sp/Yp, although additional acidic residues
are required to optimise its phosphorylation
efficiency, explaining why clusters of glutamic and
aspartic acids are found on the C-terminal side
(and sometimes also on the N-terrninal side) of most
of its physiological sites [80]. Pertinent to this are a
comment and a warning. The comment is that the
consensus sequences of CK2 and CDKs are mutually
incompatible: CDKs in fact need a proline at n+1
and select a basic residue at n+3, whereas CK2
needs an acidic residue at n+3 and selects another
acidic residue at n+ 1, where a proline actually
acts as a powerful negative determinant [81]. The
warning is that the popular rule that phospho-
rylated serines/threonines followed by multiple
acidic residues are invariably targets for CK2 or
'CK2-like' enzymes should be applied cautiously
since we know that the Golgi 'casein kinase', once
believed to be expressed only in lactating mammary
gland, is present in other tissues as well [4]. As
shown in table 1, the consensus for G-CK, S-X-E/Sp,
is similar to that for CK2, and in this case also
additional acidic residues improve the
phosphorylation efficiency [82]. Consequently
phosphoacceptor sites having acidic residues at both
n+2 and n+3 (with Glu at n+2) can a priori be
substrates of either CK2 or G-CK, this latter being by
no means a 'CK2-like' kinase.
The structural basis for the acidophilic nature of
CK2 has been studied by mutational analysis of those
basic residues that are conserved among CK2s from
different sources, but are not conserved in other
protein kinases [26, 38, 83, unpublished data]. These
studies have led to the identification of K49, K74-77,
K79/R80, H160 and R191/R195/K198 as residues
implicated in the recognition of specificity
determinants (table 2).

Table 2. CK2a residues implicated in substrate recognition.
Residue(s) Region
K49 "Gly loop" (Subd. I)
K74, K75, K76, K77 Connection subd. II-III
K79,RBO subd. III
HI60 catalytic loop (subd. VIB)
R191, R195, K198 loop p+ 1 (subd. VIII)
Regulation
It may appear frustrating that the mode of
regulation of a protein kinase endowed with a
canonical quaternary structure, equipped with two
'regulatory' subunits and able to perform both
autocatalytic and heterologous phosphorylation still
remains obscure. But this is precisely the case,
exacerbated by the fact that CK2 is one of the most
pleiotropic kinases, impinging on myriad substrates
and implicated in cell proliferation and
transformation. This situation appears even worse
compared with other kinases of the same branch,
MAP kinases, CDKs and GSK3, which all fall into the
wide category of kinases, the activity of which is
triggered by phosphorylation within their 'activation
loop' [40, 84]. In the case of MAP kinases two
residues within the loop (T183 and Y185) are
phosphorylated by dual-specificity Mek (itself a
kinase with a phosphorylatable ac·tivation loop),
while in CDKs a single threonine is phosphorylated,
T160 in CDK2 (see fig. IE). In both cases
phosphorylation is heterologous, while with other
kinases (e.g. PKA, many tyrosine kinases) an
autocatalytic phosphorylation occurs. Inspection of
the activation loop of CK2 (fig. IE) reveals that it
contains no Ser /Thr residues, although it does have
one tyrosine residue. However, despite several
attempts, no substantial phosphorylation of CK2 by
tyrosine kinases has been detected. The failure of
CK2 to fulfil the activation loop phosphorylation
rule is hard to accept since CK2 falls into the
category of 'RD kinases', in which the invariant
catalytic aspartate (DI56) is preceded by an
arginine, the neutralisation of which by the
activation segment phospho residue is required for
catalysis [40). Two exceptions are provided by
phosphorylase kinase and CKI (mammalian),
where a glutamate and an asparagine,
respectively, substitute for the phosphoresidue
and interact with the crucial arginine, ensuring
that the catalytic site has a constitutively active
conformation [40]. In CK2 a/a' the residue
homologous to CDK2 Thr-I60 is an asparagine, and a
glutamic acid is located two positions upstream. It
will be interesting to see if CK2 inactivated by
mutating these residues. However, although such a
result would account for the constitutive activity, it
would not answer the question as to Iww CK2 is
regulated.
85
CHAPTER 7/ PROTEIN KINASE CK2
Determinant recognized
n+2
n+3/n+4
n+3
n-2
n+l
A second regulatory device of CDKs is the
phosphorylation of two adjacent residues (T14-YI5)
within the 'Gly loop', which keeps the kinase silent
until a dual-specificity phosphatase removes the
phosphate. Since the phosphorylation of Tyr-15 alone
is sufficient to inactivate CDKs, it seems intriguing
that this tyrosine is conserved in CK2 (see fig. 1), but
not in the great majority of protein kinases. It is
disappointing therefore that this tyrosine cannot be
shown to be phosphorylated by tyrosine kinases.
In summary there is no evidence whatsoever that
CK2 might undergo tight control through
phosphorylation/ dephosphorylation, although it
cannot be ruled out that autophosphorylation
and/ or phosphorylation by proline-directed kinases,
shown to occur both in vitro and in vivo, might subtly
tune some properties of CK2. In particular,
autophosphorylation at the N -terminus of the ~­
subunit, occurring through an intramolecular
mechanism [85], has been shown to correlate with a
modest decrease of activity [54] in accordance with
the observation that the autophosphorylation site is
functionally related to and possibly physically
associated with the acidic cluster responsible for the
down-regulatory properties of the ~-subunit [55].
This finding could account for an early observation
[86] that CK2 is slightly, yet significantly, more active
if it is assayed in the presence of protein
phosphatases that presumably keep it in a
dephosphorylated state. In contrast it cannot account
for the plethora of reports that appeared between
1987 and the early nineties, supporting the view that
CK2 phosphorylation correlates with short term
increase in activity in response to serum, hormones
and growth factors. A compilation of references
relating to this can be found in [87], where most of
these purported effects were re-evaluated and
ultimately dismissed on the basis that no
reproducible increases of CK2 activity were observed
in response to the treatments previously described.
Another intriguing analogy between CDKs and
CK2 is provided by the cyclins, which assist CDKs
with a third mode of regulation: the activity of CDKs
is dependent upon their association with cyclins, the
. concentration of which varies throughout the cell
cycle. Abrupt proteolysis will deprive an individual
CDK of its cognate cyclin, leaving it in an inactive
L. A. PINNA AND F. MEGGIO
form until the next increase in the concentration of
eyelin (reviewed in [88, 89]). Apart from differences
in quaternary structure (dimeric vs. tetrameric), the
association of cyelins with CDKs is reminiscent of
that of the ~-subunit with the catalytic subunits of
CK2. Like eyelin, the ~-subunit is required for full
activity (at least in vitro with peptide substrates) and,
as discussed earlier (see section entitled ''Structure of
the ~-subunit"), it contains a motif similar to the
destruction box that targets proteolytic degradation
of eyelin. However, there are two strong arguments
against the hypothesis that the 'Merry Widow'
model of CDKs/cyelins may apply to CK2. These
are, first, the extremely high affinity of ~ - a
interactions, resulting in no detectable exchange
between bound ~ and the large excess of free ~­
subunits (unpublished data); and second, the
observation that the half-lives of the catalytic and ~­
subunits are quite similar [7] and, apart from special
cases, excess of the a over the ~ subunit has not been
described (rather the opposite has been reported [90,
91]).
It is still conceivable, however, that before the
tight a-~ links are formed other ligands might
interact with the a-subunit, inducing changes in
activity and possibly slowing down its assembly
with ~. Such a possibility appears strengthened by
the observation that assembly of CK2 holoenzyme in
HeLa cells nuelei, where the free subunits are
separately imported, is a slow process [7]. This study
has shown that the half-lives of the three CK2
subunits are similar (in the range of 24-29 h) and that
a chase of 10-14 h is required to reach complete
complex formation for the a2~2 form. Even if, as one
of the authors argues, these low values reflect an
exchange of subunits into the existing complex,
rather than true rates of complex assembly (Marshak,
personal communication), it must still be assumed
that a and ~. not combined to each other, are
available at certain times. It is not elear what it is
within the cell that slows down a process that is
virtually instantaneous when the recombinant a and
~-subunits are combined in vitro. It is conceivable,
however, that during these lags the a-subunit might
come in contact with ligands that hamper its
association with the ~-subunits. Pertinent to this is
the observation that the stellate protein can interact
with the a-subunit and substitute for some functions
of the ~-subunit, although not others (e.g. stimulation
by polylysine) [92]. Likewise, other known ligands of
CK2a, e.g. HSP-90 [34], nueleolin [35], PP2A [39],
could bind to the free catalytic subunit, hindering or
slowing down the assembly of the canonical
heterotetramer.
The same may apply to the free ~-subunit, many
ligands of which are being identified and in which an
NLS motif is lacking. It should be noted that
according to this hypothetical scenario
86
immunochemical methods would still detect
stoichiometric amounts of a and ~ in the same
compartment, as if canonical heterotetramers were
there. It is also likely that once extracted and
submitted to purification procedures these separated
a and ~ subunits would dissociate from their
temporary partners and give rise to the 'regular'
holoenzyme which is also much more stable to
proteolytic degradation than isolated subunits, thus
accounting for the failure to detect forms of CK2
other than the heterotetramer during purification.
This would also be consistent with the early and
puzzling observation that a relatively high salt
concentration is needed in order for CK2 to bind to
phosphocellulose, the first-choice resin for its
purification [93]. At low salt concentration CK2 does
not bind, while, once bound at 0.25 M NaCl it needs
a very high salt concentration (up to 0.7M) to elute it
[93].
Despite all these caveats, which may open new
perspectives on the regulation of CK2a before
and/or independent of association with ~-subunit,
we have to admit that it is very unlikely that the
heterotetrameric form of the holoenzyme is just an in
vitro artefact which does not exist in vivo. Thus, we
are left with two main questions: first, what is the
role of the ~-subunit? Second: if the ~-subunit is a
'regulatory' subunit, how can the activity of CK2 be
reversibly modified through it?
Until a few years ago the answer to the first
question appeared self-evident: reconstitution
experiments had repeatedly shown that the ~­
subunits were required for optimal catalytic activity.
Today, however, this dogma has become open to
question. First, it has become evident that with
certain substrates, notably with calmodulin, the
addition of the ~-subunit has a dramatically negative
effect, causing full inhibition of phosphorylation [53,
94]. Moreover, such a down-regulatory potential of ~
also operates with canonical peptide substrates,
where it is normally masked by the predominant
positive effect. This effect has been uncovered by
generating mutants (altered in the 55-64 acidic
eluster) that are hyperactivatory with peptide
substrates and no more inhibitory with calmodulin
[53]. The objection that calmodulin is unique in this
respect does not hold since a higher phosphorylation
by a as compared to holoenzyme is also observable
with NDPK (unpublished) and with MDM2 [95] and
it is to be expected that all substrates whose
phosphorylation by CK2 in vitro is almost fully
dependent on polylysine (e.g. ODC, elathrin b-chain)
will be better phosphorylated by the free a-subunit
than by the holoenzyme. Moreover, the overall effect
of the ~-subunit seems to depend on a variety of
conditions since, under special circumstances the ~­
subunit has also been shown to decrease the
phosphorylation of peptide substrates [96]. The

dogma that ~ is required merely to ensure optimal
activity of CK2 is also contradicted by the
observation that in some organisms monomeric
forms of CK2a, devoid of the ~-subunit, exist, and by
experiments showing that in S. cerevisiae in which
both a and a' genes were disrupted viability was
rescued and the normal phenotype restored by
expression of the Drosophila a subunit, which is
unable to assemble with the endogenous ~ and W
subunits [8]. The different behaviour of S. pombe,
where disruption of the ~ gene is detrimental and
causes a drop in CK2 activity [97], could be
explained by assuming that S. pombe CK2 catalytic
subunits are intrinsically poorly active and that in
this special case the p-subunit is required to achieve
sufficient activity. Consistent with this interpretation,
the p-subunit acidic cluster responsible for negative
regulation is not conserved in S. pombe, suggesting
that this p subunit is solely designed for up-
regulation. In vertebrates, as in S. cerevisiae, the a-
subunit alone is functionally autonomous, as in
transgenic mice its expression is sufficient to
generate lymphomas [11] (see section entitled "CK2
in neoplasia: a vicious circle?"). It would be
interesting to see whether p-subunit co-expressed
with a potentiates or attenuates the oncogenic
potential of the latter. This could tell us if, as far as
the substrates implicated in tumorigenesis are
concerned, the predominant function of p in vivo is to
up- or to down-regulate the activity of CK2.
With regard to the second question - can the p-
subunit mediate reversible regulation of CK2? - two
classes of cationic compounds are known to affect
the activity of CK2 by interacting with the ~-subunit,
polyamines (particularly spermine) and polybasic
peptides, e. g. polylysine and some histones. Despite
their similar stimulatory effect and their common
polycationic nature these two classes of effectors
operate in different ways as they interact with
partially different regions of the p-subunit and only
the latter are capable of triggering the
phosphorylation of calmodulin by CK2 holoenzyme.
In other words, while spermine potentiates the
positive effect of the p-subunit (being equally
effective whether p wild type or 'hyperactivatory' p
mutants are used to reconstitute the enzyme
[unpublished results», polylysine and similar
polybasic peptides reverse the negative effect of the
~-subunit. It is curious that spermine counteracts the
activatory effect of polybasic peptides in a dose-
dependent manner, despite the fact that they bind to
distinct sites. This leads to the paradoxical outcome
that, under certain circumstances, spermine can
operate as an inhibitor of the activity of CK2 [98].
The problem with these polycationic effectors is
that, on the one hand spermine, which can be
regarded as a bona fide second messenger, displays
only a collateral effect by potentiating stimulation by
87
CHAPTER 7/ PROTEIN KINASE CK2
the Ji-subunit; whereas, on the other hand, polybasic
peptides, which alone are capable of triggering all-
or-none effects like those expected of acute
regulation, can hardly be considered mediators of
short-term reversible regulation. Among the
naturally occurring basic proteins that mimic
polylysine, histones deserve attention in view of the
fact that CK2 concentrates in nuclei, where
phosphorylation of calmodulin at the same sites
affected in vitro by CK2 supplemented with
polylysine has also been shown to occur [99]. On the
other hand it should be emphasised that calmodulin
phosphorylation by CK2 holoenzyme is neither
promiscuously triggered by any kind of long basic
stretch (only some histones and protamines are
effective) [53] nor requires a large number of basic
residues, as a short peptide reproducing the basic
stretch of CK2 a between subdomains II and III
(which also includes the nuclear localisation signal)
can induce the same effect [98]. This makes it
conceivable that proteins which include suitable
basic motifs, possibly including some of the many
cellular partners of CK2~ (see below), might
overcome down-regulation by ~, triggering the
targeting of protein substrates the phosphorylation
of which is otherwise inhibited by this subunit. In
this connection a breakthrough may come from a
recent study of the phosphorylation of the MDM2
protein inhibitor of p53. This protein, the
phosphorylation of which is catalysed by the isolated
CK2a subunit more efficiently than by the
heterotetrameric holoenzyme (like in the case of
calmodulin), is stimulated by the carboxy-terminus
of p53, mimicking the effect of polylysine [95]. A
similar stimulatory effect is exerted by another
partner that interacts with p, FGF-2 [1(0). In this case,
however, it is not known whether FGF2 operates by
mimicking the effect of polylysine, although the
predominance of basic residues in it would suggest
that this may be the case.
The discovery of an increasing number of
proteins capable of interacting with the p-subunit of
CK2 opens new perspectives on the regulation of this
kinase. Although a few such 'interacting partners'
(e.g. p53) were known some years ago, in most cases
it has been the recent use of the two-hybrid system
that has paved the way towards their detection. By
applying this approach to a HeLa cell cDNA library
Boldyreff and Issinger [59] isolated more than 1000
clones, and they have calculated that 164 of these
represent strong interacting partners of CK2~. Only
some of these have so far been identified by these
workers or by other groups. However, the proteins
identified include A-Raf [62] , the ribosomal protein
L5 [101], FGF-2 [100], Nopp140 [58], ferritin heavy
chain, a-prolyl-4-hydroxylase, catepsin-C, UBE1,
HSP70, calmodulin and Raf-1 (c. Cochet and O.
Filhol, personal communication). Except in few cases
it is not known which functional consequences the
L. A. PINNA AND F. MEGGIO
Table 3. Alphabetical list of proteins phosphorylated by protein kinase CK2. Proteins without reference are referenced in (12) or (109).

Al (human hnRNP) HDAgs (Hepatitis 0 virus antigens) [134]

ABF-l (yeast) [110] HIV -1 Rev protein [135]
Adenovirus Ela HIV-1 viral protein
Androgen receptor HMG-14
c-jun HMG-l
c-Myb HMG-Iike protein PI
c-Myc hnRNP
Ca2+ channel blockers receptor HPVE7
Caldesmon Hsp90
Calmodulin Human centrom. protein B (CENP-B) [136]
Calnexin Human complement protease Clr [137]
Calreticulin (spinach) [111] Human estrogen receptor [138]
Calsequestrin IGF-II receptor
Caveolin [112] IGFBP-l
CDC37[113] IkBa [139]
CD45 tyrosine phosphatase [114] IkB~ [140]
CIathrin light b-chain Inhibitor-2 of PPI
Coactivator pIS (PC4) [115] Insulin receptor
CREB IRS-l
CREM LD lipoprotein receptor
CTP:phosphocol. cytidyltransferase [116] Lipoprotein inhibitor
DARPP-32 Mannose-6-phosphate receptor
DNA ligase MAP-IB
DNA topoisomerase I Max protein
DNA topoisomerase II MDM2 [95]
DSIP peptide Measles virus P-protein [141]
Dynamin [117] MEF2C (myocyte enhancer factor) [142]
Dystrophin MRFs (MRF4 and MyoD) [143]
E47 [118] mRNA particles
EF-l 0 [119] mUBF
EF-l~ MYCN (human neuroblastoma cell)
Egr-l (early growth response) [120] Myosin heavy chain
eIF-2a [121] (brain, smooth and skeletal muscle)
eIF-2~ Myosin light chain
eIF-3 N-myc
eIF-4B NBP-60 (NLS binding protein) [144]
eIF-5 Nef
eIF4e/p20 (5. cerevisiae) [122] Neural cell adhesion molecule L1 [145]
Engrailed protein (Drosophila) [123] NIPP-l [146]
Epstein-Barr Nuclear Antigen 2 nm23/NDPK [147]
Epstein-Barr SM P-protein [124] Nuclear DNA-binding protein p210
Epstein-Barr virus ZEBRA protein Nucleolar p130 [148]
Fibrinogen Nucleolar protein B23
Filaggrin Nucleolin
FK506 binding protein Nucleoplasmin [149]
Fragmin (Physarum polycefalum) [125] Ornithine decarboxylase
Furin [126] Osteopontin
Gap43 (neuromodulin) Po, PI and P2 ribosomal proteins
GB glycoprotein (cytomegalovirus) [127] p120 cell proHf. nucleolar protein
GBF-l transcr. factor (Arabidopsis) [128] p13 and p38 yeast ribosomal proteins [150]
Glycogen synthase p25 (Xenopus laevis) [151]
Glycophorin p34 (chloroplast ribonucleoprotein)
Glycyrrhizin [129] p34cdc2
gp96 (soybeans) [130] p38 RNA-binding protein [152]
GRP94 of sarcoplasmic reticulum [131] p53
Guanine nucleotide exchange factor p65 synaptic vesicles
HASPP28 (rat brain) [132] p98 (sea urchin egg)
HCP (hist.-rich Ca++ bind. protein) [133] Pea lamin like protein

88

PHAS-1 [153]
PKA R-II subunit
PKC
Platelet Va and VIlla factors
PP2C
pp35 proliferation nuclear P-protein
pp65 (human cytomegalovirus) [154]
Progesterone receptor [155]
Proteasome (30 kDa)
Proteasome complex (CB and C9) (156)
Prothymosin a
Pseudorabies virus pp-62
PU-1 transcription factor
RAB-17
Rat epididymal fat cell p22
Respiratory syncytial virus P-protein [157)
RlPP
RlPP-I and -II phosphophoryns (158)
RNA polymerase I
RNA polymerase II
RNA polymerase III (159)
sarcalumenin (133)
Serum response factor
Spectrin
binding of these 'partners' might have. It is to be
expected, however, that at least some of these
interacting molecules will impinge on the regulatory
properties of the ~-subunit, ultimately altering the
catalytic activity of CK2, as this appears to be so in
the case of p53 (102).
Another way in which CK2 might be regulated is
suggested by the observation that under certain
circumstances the tetrameric holoenzyme tends to
aggregate giving rise to high molecular weight
polymers [103-105). Depending on the salt
concentration, three kinds of supramolecular forms
of CK2 have been detected, 'rings', 'thin' filaments,
and 'thick' filaments [105]. It has been suggested that
physiological salt concentration and saturating
substrate concentration would favour tetrameric
rings ([a2~2)4), rather than the canonical holoenzyme
(105). The actual occurrence of these aggregated
forms of CK2 within the intact cell and their possible
physiological relevance remain to be established.
As a final point one should remember that
regulation of constitutively active protein kinases
refractory to any other kind of control can be
eventually afforded by protein phosphatases
impinging on their phosphorylatable substrates.
Mutatis mutandis, a pertinent example could be that
of cyclins, the changes of intercellular concentration
of which are crucially dependent on their proteolytic
breakdown [89). Although it involves considerations
of cellular energy expenditure, it is plausible that, by
analogy, the phosphorylation state of some or all of
the targets of CK2 may depend on oscillations in
protein phosphatase activity while the activity of the
89
CHAPTER 7/ PROTEIN KINASE CK2
Sperrnin binding protein
Spi-B/Spi-1 hematop. transcr. fact. (160)
Stathrnin [161]
SV40 large T antigen
Synaptotagrnin
T-substrate (wheat germ)
Tal-1 [162)
Tau protein (163)
TFIIA coactivator pIS (164)
TRa2 (thyroid hormone receptor) (165)
Troponin-T
Tryptophanyl-tRNA synthetase [166)
Tubulin
VAMP / synaptobrevin [167)
Viral P-protein
Vitamin D receptor
VP1 (polyoma virus capsid) [168)
VpU (HIV-l) [169]
VSV (ves. stomatitis virus) P-protein
VZV (varicella zoster virus) protein
XFG-5-1 (Xenopus zinc finger) protein [170]
xNopp180 (Xenopus laevis) [171)
Yeast SRP-40 (homologue to Nopp140) [172)
ZFPs (Xenopus laevis) (173)
kinase remains unchanged. As discussed elsewhere
[106), the acidic nature of phosphoacceptor sites
affected by CK2 suggests that the phosphatase
counterpart of CK2 must be PP2A, as it is the only
Ser /Thr phosphatase that tolerates acidic residues on
the C-terminal side of the phosphoaminoacid [106). It
may be worth noting in this connection that there are
some intriguing analogies between PP2A and CK2.
PP2A has a constitutive regulatory subunit which,
like CK2~, does not exert acute control of catalytic
activity, although its modulation seems to be
mediated by a variety of interacting proteins [107,
108). This is reminiscent of the 'interacting partners'
of CK2 ~ discussed above. In the light of this it
appears even more intriguing that PP2A is itself a
partner of CK2a [39].
SUBSTRATES: AN UPDATE
One of the paradoxes in the CK2 story is that,
after having been a kinase without substrates for two
decades (the only ones known being foreign: casein
and phosvitin), it suddenly turned out to be one of
the most pleiotropic kinases known [9). The list of
substrates of CK2, which in 1980 included just three
proteins, had increased to 35 proteins by 1990 [12),
and to 99 proteins by 1993 (109). Table 3 presents a
list of 162 proteins that were known to be substrates
of CK2 at the end of 1996, although there is no
pretension that this is complete.
It is possible that in a few of these cases the
identification of CK2 as the physiological
phosphorylation agent is not incontrovertible. Often
identification is based on the finding of a clear-cut
L. A. PINNA AND F. MEGGIO
CK2 signature sequence surrounding the site of
phosphorylation in vivo, corroborated by the parallel
demonstration that the same site is phosphorylated
by CK2 in vitro . As outlined above (see section
entitled "Catalytic properties and specificity") this
might cause mistaken inferences for G-CK because of
the partially overlapping site-specificity of G-CK and
CK2 (see also table 1). This caveat, however, is
limited to a minority of proteins in table 3, e.g. furin
and the mannose-6-phosphate receptor, which are
associated with the Golgi apparatus where G-CK
predominates over CK2, whereas in other subcellular
compartments G-CK is virtually absent [4].
Even more striking than the number, is the nature
of the substrates listed in table 3, many of which are
implicated in a wide variety of cellular functions
(reviewed in [1]). A common denominator of the
majority of substrates of CK2 is their participation in
signal transduction pathways or in biochemical
events triggered by those signals. Especially relevant
to this review are the many proteins in table 3 that
playa role in gene expression, protein synthesis, cell
division and proliferation. Many of these are also
targets for CDKs, although, as discussed above (see
section entitled "Catalytic properties and specificity")
the CK2 and CDKs sites are rigorously differentiated
by mutually incompatible consensus sequences.
REQUIREMENT IN CELL DIVISION CYCLE
It has been emphasised from the very beginning
of this review that CK2 is essential for viability. This
was unambiguously shown in S. cerevisiae, where
disruption of CK2 genes is lethal [8]. Genetic
manipulations in Dictyostelium discoideum [20] and S.
pombe [174] support the same conclusion. An
important outcome of the study on S. cerevisiae,
which implicitly pointed to cell division as the event
for which CK2 may be indispensable, was the
phenotype of the defective cells, which continued to
grow after cell cycle progression was arrested, and
retained significant capacity for protein and DNA
synthesis [8]. This argues against the possibility that
cell cycle arrest caused by disruption of the CK2 gene
is an indirect consequence of a general decline in
metabolic activity and/or biosynthetic competence.
Moreover, some of the cells arrested by CK2
depletion exhibited an elongated bud, a phenotype
shared by several cell division cycle mutants [175].
The implication of CK2 in cell division would also
be consistent with a variety of additional
observations. Some of these are merely
circumstantial: e.g. the phylogenetic relatedness of
CK2 and the cell division cycle kinases; the ability of
CK2 to phosphorylate the cell division cycle 'engine',
p34cdc2 [176] and to be phosphorylated on both its a
and ~-subunits by CDKs [49, 63] (although these
phosphorylations do not seem to have any effect on
90
activity); phosph~lation-independent activation of
CK2 a by the p34 c2 / cyclinB complex, in vitro [177];
and inclusion in the list of CK2 substrates (table 3) of
many proteins known to playa role in cell division
and proliferation. Other arguments however are
more pertinent and stringent, such as the observation
that micro injection of active CK2 into Xenopus
oocytes accelerates meiotic maturation induced by
mitosis-promoting factor [178], and the inhibition of
serum-induced cell cycle progression by
microinjection of antibodies against the ~-subunit of
CK2 into Go-synchronised human fibroblasts [179].
Incidentally this latter result supports the view that
the competent form of CK2 in this case is the
canonical tetramer including the ~-subunit, and not
the free catalytic subunits.
Additional incontrovertible evidence for a cause-
effect linkage between CK2 activity and cell cycle
progression in S. cerevisiae was provided by the
Glover group which generated five independent
temperature-sensitive alleles of one of the genes
encoding the CK2 catalytic subunits (CKA2) [113].
One strain arrested within a single cell cycle, and the
analysis of the arrested cell population showed that
they were in Gt. The conclusion that CK2 is required
in Gt was confirmed by flow-cytometry of
hydroxyurea-synchronised cells, which, in addition,
indicated that CK2 is also required for cycle
progression in G2 and/or mitosis. A requirement for
CK2 during Gt is consistent with data from
mammalian systems, notably the experiments
involving microinjection of anti-CK2~ antibodies, in
which cells were inhibited at multiple points in Gt
[179], and reports of increased concentrations of CK2
just before the GtiS transition [180, 181]. On the
other hand a function of CK2 downstream of mitotic
CDK activity (supported by its requirement in
G2/M) would be consistent with the dramatic
increase of phosphorylation of CK2 a and ~ subunits
at CDK target sites found in animal cells arrested in
mitosis [49].
Although the targets of CK2 during the cell cycle
are still unknown, Hanna et al. [113] speculate that,
besides cdc28 (the S. cerevisiae homologue of p34cdc2)
itself, other potential candidates in Gt could be cdc37
and DNA ligase I, both substrates of CK2 (see table
3). In G2/M they suggest the prime candidate to be
topoisomerase II, for which physiological targeting
by CK2 is well documented [182], and the
implication of which in CK2-mediated functions is
supported by a number of coincidental observations
[113].
Another plausible pathway by which CK2 could
affect the cell division cycle is more indirect: through
inhibitor-2 (1-2) of protein phosphatase-I. Activation
of 1-2 requires its preliminary phosphorylation by
CK2 at seryl sites which potentiate secondary

phosphorylation of regulatory Thr-72 by GSK3 [107,
183, 184]. During mitosis 1-2 activity increases [185],
affecting PP1, the activity of which consequently
declines [88]. This prevents dephosphorylation (and
inactivation) of the dual phosphatase cdc25 which, in
consequence, can activate cdc2 by dephosphorylating
its down-regulatory T14/Y15 site (see above).
Through this cascade of events CK2 would
ultimately trigger the activation of cdc2 and drive on
mitosis.
The demonstration that CK2 is required for
progression through the cell division cycle, although
the mechanistic rationale for this requirement is still
unknown, accounts for many reports showing that
CK2 invariably rises in proliferating cells. This
general rule applies during embryogenesis [186-190]
and larval development [191]. These studies showed
that not only does CK2 tend to be higher on average
during fetal development than in the adult, but it
also undergoes cyclic oscillation and tissue-specific
variation [192]. Other 'normal' proliferating tissues
in which similar increases of CK2 have been
observed include regenerating liver [193, 194] and
intestinal colonic mucosa, where the very telling
observation was made that CK2 is particularly
elevated in the cell layer at the base of the crypts
which also is characterised by an especially high
proliferative activity [195, 196]. In the light of these
and other observations the proposal that CK2 should
be considered a 'proliferation marker' [197] still
holds.
The elevated activity of CK2 found in the basal
cell layer of the colon was accounted for by a high
level of expressed CK2, as documented by
immunochemical methods [196]. This is likely to
apply to all proliferating tissues and would be
consistent with the requirement of CK2 for the
progression of the cell division cycle, as discussed
above. However, it is also possible that CK2
expressed in proliferating tissues might have
distinctive features as suggested by altered kinetic
constants of CK2 from fetal liver [187] and by the
intriguing observation that a third type of a subunit,
termed a3, smaller than both a and a', and possibly
proteolytically generated from these, is abundant in
CK2 isolated from the nuclei of regenerating liver
[194]. Also pertinent to this may be the finding that
treatment of murine fibroblasts with serum
specifically evokes the appearance of a' mRNA
which is already evident two hours after treatment
(Oliviero, personal communication)
CK2 IN NEOPLASIA: A VICIOUS CIRCLE?
There is no doubt that proliferation and
transformation are quite distinct events, as different,
grosso modo, as health and illness. However, it is also
undeniable that cell proliferation invariably
91
CHAPTER 7/ PROTEIN KINASE CK2
accompanies the onset of tumors. It is therefore no
surprise that CK2 is elevated, not only in 'normal'
cell proliferation, but also in neoplastic tissues. This
is documented by an impressive number of reports
[10] dealing with a wide variety of pathological
situations, including hepatoma, leukemic cells,
benign prostatic hyperplasia, immortalised cell lines
and several solid tumors such as colorectal, bladder,
kidney and mammary carcinomas. This rises the
crucial question as to whether the increase of CK2
observed in all these cases is just a consequence of
cell proliferation or may have a causative effect on
the onset of at least some types of tumors.
Some observations argue in favour of a
relationship between the extent of transformation
and the increase in CK2.Thus, the mean activity of
CK2 increases on passing from colorectal mucosa to
adenomas and from adenomas to carcinomas [198],
suggesting a gradual enhancement with stage,
according to the concept that colorectal carcinomas
arise from adenomas. Likewise a statistically
significant correlation between CK2 and the 'grade'
of tumors was found on analysing kidney and
bladder carcinomas [199, 200]. On the other hand the
somewhat paradoxical situation of prostate has been
reported, where the increase of nuclear CK2 is much
more dramatic in benign hyperplasia than in
carcinomas [201]. This is not entirely surprising
however if it is considered that prostatic carcinomas
do not seem to develop from hyperplastic tissue.
What is definitely striking is the general validity
of the 'CK2 increase' rule in tumors of totally
different origins. For example, CK2 is elevated in
human leukemic cells [202], in virus-transformed
murine lymphocytes [203], and even in bovine T-cells
transformed by the parasite protozoan Theileria paroa
which causes a leukemia-like disease of cattle [204].
While this could be interpreted to suggest that the
increase in CK2 activity is a consequence, rather than
a cause of lymphoid transformation, a causative
effect of CK2 on the onset of leukemia has been
demonstrated by the over-expression of CK2 in T
cells transformed by T. parva. To investigate the
possible role of CK2 in the pathogenesis of
lymphoproliferative disorders, the a-subunit of CK2
was expressed in the lymphocytes of transgenic
mice, giving rise to a stochastic propensity to
develop lymphomas [11]. Even more interesting,
such a moderate tumorigenicity was dramatically
enhanced by mating CK2 a with c-Myc transgenic
mice. This gave rise to bi-transgenic offspring with a
marked increase in the incidence of
lymphoproliferative disease [11]. The oncogenic
potential of the catalytic subunit of CK2 was
subsequently confirmed using another experimental
model taking advantage of a transgenic mouse line in
which mis-expression in thymus of tal-l (the product
of the TAL-l gene the ectopic activation of which
1. A. PINNA AND F. MEGGIO
occurs in human T-cell acute lymphoblastic
leukemia) causes leukemogenesis [205]. This was
dramatically accelerated by transgenic coexpression
of CK2 a, providing a second example of a situation
in which CK2 a has a synergistic effect in causing
lymphomas. A third example has been provided
by experiments that demonstrate quite a dramatic
acceleration of lymphoma by CK2a in p53
heterozygous and homozygous knock-out mice
(D. C. Seldin, personal communication).
Since both c-Myc and tal-1 are substrates of CK2
(see table 3) it would be tempting to suggest that the
oncogenic potential of CK2 is mediated through its
ability to phosphorylate and hyper-activate
transcription factors. This seems not to be the case,
however, at least with tal-I, the phosphorylation of
which was not significantly increased in the tal-
l/CK2a leukemic cell lines. This finding, in
conjunction with the failure of attempts to co-
immunoprecipitate tal-1 and CK2 or to observe any
further up-regulation of tal-I mRNA levels in the tal-
l/CK2a bitransgenic tumor cells, argues against a
direct effect of CK2 on tal-1 activity or expression,
supporting, rather, an indirect synergism brought
about by two independent cascades that may
converge at definite point(s).
On the basis of two well-documented premises,
i.e. that the activity of CK2 increases in dividing cells
and that a variety of proteins implicated in gene
expression are among its targets, an attractive
hypothesis would be that CK2 is implicated in a
'vicious circle' that could operate in a wide variety of
tumors, irrespective of their origin. It is conceivable,
in fact, that the physiologically high level of CK2
required during cell proliferation could become
oncogenic if other parameters susceptible to CK2
activity and with the potential to promote
transformation are qualitatively or quantitatively
altered. Given that mitogenic stimuli invariably
dictate CK2 expression even under normal
conditions, it is possible that under special (aberrant)
circumstances the action of these stimuli, which per se
are weakly oncogenic, is in turn protracted and/or
potentiated by CK2 itself, giving rise, through a
reciprocal feedback effect to the amplification of the
level and/or duration of the wave of CK2 expression.
This would increase the probability that conditions
for the onset of neoplasia are fulfilled. In other
words, the ubiquity of CK2 and its 'house-keeping'
functions - in conjunction with its remarkable
pleiotropy - could render this kinase potentially
dangerous whenever there is a disturbance of the
delicate balance between cell death and proliferation
(e.g. over-expression of tal-lor suppression of p53,
see above). This scenario could reconcile the
observation that the activity of CK2 is elevated, not
only in all tumors examined - irrespective of their
origin - but also in 'normal' proliferating tissues,
92
with the finding that, at least in some cases, such an
increase in CK2 is the definite cause, rather than just
a consequence, of tumor onset. The fact that
potentially oncogenic agents/events, whatever their
mode of operation, need to induce a mitogenic
response implies that CK2 will always be involved
in, and in some cases could be critical for, reaching a
'threshold of disregulation' leading to neoplasia.
If this hypothesis is correct, CK2 will become an
attractive target for potential anti-neoplastic therapy
because of its implication, as a contributing, yet
critically causative, agent in a wide spectrum of
tumors. More important, because its complete
inhibition would neither be required nor desirable,
an attenuation of the activity of CK2 might be
sufficient to suppress its oncogenic potential. In this
connection it is pertinent to note that in lymphomas
promoted by transgenic expression of CK2 in mice
[11] the excess of CK2a mRNA over that in non-
oncogenic situations was less than 20%.
ACKNOWLEDGEMENTS
We thank Drs. Olaf-G. Issinger (Odense,
Denmark) and David P. Leader (Glasgow, Scotland)
for reviewing the manuscript. This work was
supported by grants from the Commission of the
European Communities (Biomed 2), from Ministero
dell'Universita e della Ricerca Scientifica e Tecnologica,
from Consiglio Nazionale delle Ricerche (grant
95.02920.CT14 and Applicazioni Cliniche della
Ricerca Oncologica), from the Italian Association for
Cancer Research and from the Ministero della Sanita
(Progetto AIDS, Istituto Superiore di Sanita).
REFERENCES
1. Allende, J. E. and Allende, C. C. (1995) FASEB
J. 9, 313-323.
2. Tuazon, P.T. and Traugh, J.A. (1991) In Adv.
Sec. Mess. Phos. Res. 23,123-164.
3. Moore, A., Boulton, A.P., Herd, H.W., Jarash,
E.D. and Craig, R.K. (1985) Eur. J. Biochem.152,
729-737.
4. Lasa, M., Marin, O. and Pinna, L.A. (1997) Eur.
J. Biochem. 243, 719-725.
5. Ahmed, K., Yenice, S., Davis, A.T. and Goueli,
S.A. (1993) Proc. Natl. Acad. Sci. USA 90,4426-
4430.
6. Krek, W., Maridor, G. and Nigg, E.A. (1992) J.
Cell. Bioi. 116,43-55.
7. Chester, N., Yu, I.-J. and Marshak, D.R. (1995)
J. Bioi. Chem. 270, 7501-7514.
8. Padmanabha, R., Chen-Wu, J.L.-P. and Glover,
C.V.c. (1990) Mol. Cell. Bioi. 10,4089-4099.
9. Pinna, L.A. (1994) Cell. Mol. Bioi. Res. 40, 383-
390.
10. Issinger, O.-G. (1993) Pharmacol. Therapeut. 59,
1-30.

11. Seldin, D.e. and Leder, P. (1995) Science 267,
894-897.
12. Pinna, L. A. (1990) Biochim. Biophys. Acta 1054,
267-284.
13. Litchfield, D.W. and Liischer, B. (1993) Mol.
Cell. Biochem. 127/128, 187-199.
14. Pyerin, W. (1994) In Protein Phosphorylation
(Marks, F. ed.), pp. 117-147, VCH Weinheim,
New York, Basel, Cambridge, Tokyo.
15. Ralph, R.K., Darkin-Rattray, S. and Schofield,
P. (1990) BioEssays 12, 121-124.
16. Filhol, 0., Cochet, c., Loue-Mackenbach, P.
and Chambaz, E.M. (1994) Biochem. Biophys.
Res. Commun. 19S, 660-665.
17. Bidwai, A.P., Reed, J.e. and Glover, e.V.e.
(1994)Arch. Biochem. Biophys. 309, 348-355.
18. Collinge, M.A. and Walker, J.e. (1994) Plant
Mol. Bioi. 25, 649-658.
18a. Glover, C.V.c. In Progress in Nucleic Acid
Research and Molecular Biology, in press.
19. Dobrowolska, G., Meggio, F., Szczegielniak, J.,
Muszynska, G. and Pinna, L.A. (1992) Eur. J.
Biochem. 204, 299-303.
20. Kikkawa, U., Mann, S.K.O., Firtel, R.A. and
Hunter, T. (1992) Mol. Cell. BioI. 12,5711-5723.
21. Stigare, J., Buddelmeijer, N., Pigon, A. and
Egyhazi, E. (1993) Mol. Cell. Biochem. 129,77-
85.
22. Heller-Harrison, R.A. and Czech, M.P. (1991) J.
BioI. Chem. 266, 14435-14439.
23. Hanks, S. K. and Quinn, A. M. (1991) Methods
Enzymol. 200, 38-62.
24. Bossmeyer, D., Engh, R.A., Kinzel, V., Postingl,
H. and Huber, R. (1993) EMBO J. 12, 849-859.
25. Hanks, S.K. and Hunter, T. (1995) FASEB J. 9,
576-596.
26. Sarno,S., Vaglio, P., Meggio, F., Issinger, O.-G.
and Pinna, L.A. (1996) J. BioI. Chem. 271, 10595-
10601.
27. Owen, D.J., Noble, M.E., Garman, E.F.,
Papageorgiou, A.e. and Johnson, L.H. (1995)
Structure 3, 467-482.
28. ole-MoiYoi, O.K., Sugimoto, e., Conrad, P.A.
and Macklin, M.D. (1992) Biochemistry 31, 6193-
6202.
29. Songyong, Z., Lu, K.P., Kwon, Y.T., Tsai, L.-H.,
Filhol, 0., Cochet, e., Brickey, D.A., Soderling,
T.R., Bartleson, e., Graves, D.J., DeMaggio,
A.J., Hoekstra, M.F., Blenis, J., Hunter, T. and
Cantley, L.c. (1996) Mol. Cell. BioI. 16, 6486-
6493.
30. Hu, E. and Rubin, e.S. (1990) J. BioI. Chem. 265,
20609-20615.
31. Gatica, M., Jedlicki, A., Allende, c.c. and
Allende, J.E. (1994) FEBS Lett. 339, 93-96.
32. Vaglio, P., Sarno,S., Marin, 0., Meggio, F.,
Issinger, D.-G. and Pinna, L.A. (1996) FEBS
Lett. 380, 25-28.
93
CHAPTER 7/ PROTEIN KINASE CK2
33. Jeffrey, P.D., Russo, A.A., Polyak, K., Gibbs, E.,
Hurwitz, J., Massague, J. and Pavletich, N.P.
(1995) Nature 376" 313-320.
34. Miyata, Y. and Yahara, I. (1995) Biochemistry
34,8123-8129.
35. Li, D., Dobrowolska, G. and Krebs, E.G. (1996)
J. BioI. Chem. 271, 15662-15668.
36. Nigg, E.A., Bauerle, P.A. and Luhrmann, R.
(1991) Cell 66, 15-22.
37. Pinna, L.A. and Ruzzene, M. (1996) Biochim.
Biophys. Acta 1314, 191-225.
38. Dobrowolska, G., Meggio, F., Marin, 0.,
Lozeman, F.J., Li, D., Pinna, L.A. and Krebs,
E.G. (1994) FEBS Lett. 355,237-241.
39. Heriche, J.-K., Lebrin, F., Rabilloud, T., Leroy,
D., Chambaz, E.M. and Goldberg, Y. (1997)
Science, in press.
40. Johnson, L.N., Noble, M.E. and Owen, D.J.
(1996) CeIlS5, 149-158.
41. De Bondt, H.L., Rosenblatt, J., Jancarik, J.,
Jones, H.D., Morgan, D.O. and Kim, S.-H.
(1993) Nature 363, 595-602.
42. Knighton, D.R., Zheng, J.., Ten Eyck, L.F.,
Ashford, V.A., Xuong, N.-H., Taylor, 5.5. and
Sowadski, J.M. (1991) Science 253, 407-414.
43. Songyang, Z., Blechner, 5., Hoagland, N.,
Hoekstra, M.F., Piwnica, W.H. and Cantley,
L.e. (1994) Curro Bioi. 4, 973-982.
44. Zhang, F., Strand, A., Robbins, D., Cobb, M.H.
and Goldsmith, E.J. (1994) Nature 367, 704-711.
45. Marcote, M. J., Knighton, D.R., Basi, G.,
Sowadski, J.M., Brambilla, P., Draetta, G. and
Taylor, 5.5. (1993) Mol. Cell. BioI. 13,5122-5131.
46. Tiganis, T., House, e.M., Teh, T. and Kemp,
B.E. (1995) Eur. J. Biochem. 229, 703-709.
47. Ducommun, B., Brambilla, P. and Draetta, G.
(1991) Mol. Cell. BioI. 11,6177-6184.
48. Marin, 0., Meggio, F., Sarno,S., Andretta, M.
and Pinna, L.A. (1994) Eur. J. Biochem. 223, 647-
653.
49. Litchfield, D.W., Luscher, B., Lozeman, P.J.,
Eisenman, R.N. and Krebs, E.G. (1992) J. BioI.
Chem. 267, 13943-13951.
50. Antonelli, M., Daniotti, J.L., Rojo, D., Allende,
e.e. and Allende, J.E. (1996) Eur. J. Biochem.
241,272-279.
51. Bidwai, A.P., Reed, J.e. and Glover, e.V.e.
(1995) J. BioI. Chem. 270, 10395-10404.
52. Boldyreff, B., Meggio, F., Pinna, L. A. and
Issinger, O.-G. (1993) Biochemistry 32, 12672-
12677
53. Meggio, F., Boldyreff, B., Issinger, O.-G. and
Pinna, L. A. (1994) Biochemistry 33, 4336-4342
54. Lin, W.-J., Sheu. G.-T. and Traugh, J.A. (1994)
Biochemistry 33,6998-7004.
55. Boldyreff, B., Meggio, F., Pinna, L. A. and
Issinger, D.-G. (1994) J. Bioi. Chem. 269,4827-
4832
L. A. PINNA AND F. MEGGIO
56. Leroy, D., Filhol, 0., Delcros, J.G., Pares,S.,
Chambaz, E.M. and Cochet, C. (1997)
Biochemistry 36,1242-1250.
57. Leroy, D., Schmid, N., Behr, I.-P., Filhol, 0.,
Pares, S., Garin, J., Bourgarit, J.-J., Chambaz,
E.M. and Cochet, C. (1995) J. Bioi. Chem. 270,
17400-17406.
58. li, D., Meier, T., Dobrowolska, G. and Krebs,
E.G. (1997) ,. Bioi. Chem., in press.
59. Boldyreff, B., Mietens, U. and Issinger, O.-G.
(1996) FEBS Lett. 379, 153-156
60. Marin, 0., Meggio, F., Sarno, S. and Pinna,
L.A. (1997) Biochemistry, in press.
61. Krehan, A., Lorenz, P., Plana-Coli, M. and
Pyerin, W. (1996) Biochemistry 35,4966-4975.
62. Boldyreff, B. and Issinger, O.-G. (1997) FEBS
Lett. 403, 197-199.
63. litchfield, D.W., Lozeman, F.J., Cicirelli, M.F.,
Harrylock, M., Ericsson, L.H., Piening, c.J. and
Krebs, E.G. (1991) ,. Bioi. Chem.266, 20380-
20389.
64. Meggio, F., Boldyreff, B., Issinger, O.-G. and
Pinna, L.A. (1993) Biochim. Biophys. Acta 1164,
223-225.
65. Vallre, B.L. and Auld, 0.5. (1990) Biochemistry
29,5647-5659.
66. Gietz, D., Graham, K. C. and litchfield, D. W.
(1995) ,. Bioi. Chem. 270, 13017-13021
67. Kusk, M., Bendixen, c., Duno, M.
Westergaard, o. and Thomsen, B. (1995) ,. Mol.
Bioi. 253, 703-711
68. Pinna, L.A., Meggio, F., Marin, 0., Perich, J.W.,
Boldyreff, B. and Issinger, O.-G. (1992) In
Adenine Nucleotides in Cellular Energy Transfer
and Signal Transduction (Papa, S., Azzi, A. and
Tager, J.M. eds.) pp. 269-280, Birkhiiuser
Verlag, Basel, Switzerland.
69. Issinger, O.-G., Brockel, c., Boldyreff, B. and
Pelton, J. T. (1992) Biochemistry 31, 6098-6103.
70. Jakobi, R. and Traugh, J.A. (1995) Eur. J.
Biochem.230,1111-1117.
71. Meggio, F., Boldyreff, B., Marin, 0., Pinna,
L.A. and Issinger, O.-G. (1992) Eur. J. Biochem.
204,293-297.
72. Whitehouse, S., Feramisco, J.R., Casnellie, J.E.,
Krebs, E.G. and Walsh, D.A. (1983) ,. Bioi.
Chem. 258, 3693-3701.
73. Cole, P.A., Bum, P., Takacs, B. and Walsh, C.T.
(1994) J. Bioi. Chem. 269,30880-30887.
74. Meggio, F., Donella Deana, A., Ruzzene, M.,
Brunati, A.M., Cesaro, L., Guerra, B., Meyer,
T., Mett, H., Fabbro, D., Furet, P.,
Dobrowolska, G. and Pinna, L.A. (1995) Eur. J.
Biochem. 234, 317-322.
75. Meggio, F., Shugar, D. and Pinna, L.A. (1990)
Eur. ,. Biochem. 187,89-94.
76. Dobrowolska, G., Muszynska, G. and Shugar,
D. (1991) Biochim. Biophys. Acta 1080, 221-226.
94
77. Szyszka, R., Grankowski, N., Felczak, K. and
Shugar, D. (1995) Biochem. Biophys. Res.
Commun. 208,418-424.
78. Vesely, J., Havlicek, L., Strnad, M., Blow, J.J.,
Donella-Deana, A., Pinna, L.A., Letham, D.S.,
Kato, J.Y., Detivaud, L., Leclerc, S. and Meijer,
L. (1994) Eur. ,. Biochem. 224, 771-786.
79. Meijer, L., Borgne, A., Mulner, 0., Chong,
J.P.J., Blow, J.J., Inagaki, N., Inagaki, M.,
Delcros, J.G. and Moulinoux, J.P. (1997) Eur. ,.
Biochem. 243, 527-536.
SO. Meggio, F., Marin, O. and Pinna, L.A. (1994)
Cell. Mol. BioI. Res. 40,401-409.
81. Marin, 0., Meggio, F., Draetta, G. and Pinna,
L.A. (1992) FEBS Lett. 301, 111-114.
82. Lasa-Benito, M., Marin, 0., Meggio, F. and
Pinna, L.A. (1996) FEBS Lett. 382, 149-152.
83. Sarno, S., Boldyreff, B., Marin, 0., Guerra, B.,
Meggio, F., Issinger, O.-G. and Pinna, L.A.
(1995) Biochem. Biophys. Res. Commun. 206,171-
179.
84. Smith, J.L., Silveira, L.A. and Spudich, J.A.
(1996) EMBO ,.15, 6075-6083.
85. Meggio, F., and Pinna, L.A. (1984) Eur. ,.
Biochem. 145, 593-599.
86. Agostinis, P., Goris, J., Pinna, L.A. and
Merlevede, W. (1992) Biochem J. 248, 785-789.
87. litchfield, D.W., Dobrowolska, G. and Krebs,
E.G. (1994) Cell. Mol. BioI. Res. 40, 373-381.
88. King, R.W., Jackson, P.K. and Kirschner, M.W.
(1994) Cell 79, 563-571.
89. Udvardy, A. (1996) Eur. J. Biochem. 240, 307-
313.
90. Stalter, G., Siemer,S., Ziegler, M., Remberger,
K. and Issinger, O.-G. (1994) Biochem. Biophys.
Res. Commun. 202, 141-147.
91. Liischer, B. and litchfield, D.W. (1994) Eur. J.
Biochem. 220, 521-526.
92. Bozzetti, M.P., Massari, S., Finelli, P., Meggio,
F., Pinna, L.A., Boldyreff. B., Issinger, O.-G.,
Palumbo, G., Ciriaco, c., Bonaccorsi, S. and
Pimpinelli, S. (1995) Proc. Nat!. Acad. Sci. USA
92,6067-6071.
93. Baggio, B., Pinna, L.A., Moret, V. and
Siliprandi, N. (1970) Biochim. Biophys. Acta 212,
515-517.
94. Meggio, F., Boldyreff, B., Marin, 0., Marchiori,
F., Perich, J. W., Issinger, O.-G. and Pinna, L.
A. (1992) Eur. J. Biochem. 205, 939-945
95. Guerra, B., Goetz, c., Wagner, P. Montenarh,
M and Issinger, O.-G. (1997) Oncogene" in
press.
96. Tiganis, T., House, C.M. and Kemp, B.E. (1993)
Biochim. Biophys. Acta 1203, 282-289.
97. Roussou, I. and Draetta, G. (1994) Mol. Cell.
Bioi. 14,576-586.
98. Sarno,S., Marin, 0., Meggio, F. and Pinna,
L.A. (1993) Biochem. Biophys. Res. Commun. 194,
83-90.

99. Quadroni, M., James, P. and Carafoli, E. (1994)
J. BioI. Chem. 269, 16116-16122.
100. Bonnet, H., Filhol, 0., Truchet, I., Brethenou,
P., Cochet, c., Amalric, F. and Bouche, G.
(19%) J. BioI. Chern. 271, 24781-24787.
101. Kim, J.-M., Cha, J.-Y., Marshak, D.R. and Bae,
Y.-S. (1996) Biochem. Biophys. Res. Commun.
226, 180-186.
102. Filhol, 0., Baudier, J., Delphin, c., Loue-
Mackenbach, P., Chambaz, E.M. and Cochet,
C. (1992) J. BioI. Chern. 267, 20577-20583.
103. Meggio, F., Brunati, A.M. and Pinna, L.A.
(1983) FEBS Lett. 160,203-208.
104. Glover, C.V.c. (1986) J. BioI. Chern. 261, 14349-
14354.
105. Valero, E., De Bonis, S., Filhol, 0., Wade, R.H.,
Langowski, J., Chambaz, E.M. and Cochet, C.
(1995) J. BioI. Chern. 270, 8345-8352.
106. Pinna, L.A. and Donella-Deana, A. (1994)
Biochim. Biophys. Acta 1222,415-431.
107. Cohen, P. (1989) Annu. Rev. Biochem. 58,453-
508.
108. Shenolikar, S. (1994) Annu. Rev. Cell BioI. 10,
55-86.
109. Pinna, L.A., Meggio, F. and Sarno, S. (1995) In
Biochemistry of Cell Membranes (Papa, S. and
Tager, J.M. eds.) pp. 15-27, Birkiiuser Verlag,
Basel, Switzerland.
110. Upton, T., Wiltshire, S., Francesconi, S. and
Eisenberg, S. (1995) J. BioI. Chem.270, 16153-
16159.
111. Baldan, B., Navazio, L., Friso, A., Mariani, P
and Meggio, F. (1996) Biochern. Biophys. Res.
Commun. 231,498-502.
112. Sargiacomo, M., Scherer, P.E., Tang, Z.L.,
Casanova, J.E. and Lisanti, M.P. (1994)
Oncogene 9, 2589-2595.
113. Hanna, D.E., Rethinasvamy, A. and Glover,
C.V.c. (1995) J. BioI. Chern. 270,25905-25914.
114. Stover, D. R. and Walsh, K. A. (1994) Mol. Cell.
Bioi. 14,5523-5532.
115. Kaiser, K., Stelzer, G. and Meisterernst, M.
(1995) EMBO J. 14,3520-3527.
116. Wieprecht, M., Wieder, T., Paul, c., Geilen, C.
C. and Orfano, C. E. (1996) J. BioI. Chem. 271,
9955-9961.
117. Robinson, P.J., Sontag, J.M., Liu, J.P., Fykse,
E.M., Slaughter, c., McMahon, H. and Sudhof,
T.C. (1993) Nature 365,163-166.
118. Johnson, S. E., Wang, X., Hardy, S.,
Taparowsky, E. J. and Konieczny, S. F. (1996)
Mol. Cell. BioI. 16, 1604-1613.
119. Minella, 0., Cormier, P., Morales, J., Poulhe,
R., Belle, R. and Mulner-Lorillon, O. (1994)
Cell. Mol. BioI. Noisy -Ie-grand 40, 521-525.
120. Jain, N., Mahendran, R., Philp, R., Guy, G. R.,
Tan, Y. H. and Cao, X. (1996) J. BioI. Chern. 271,
13530-13536.
121. Feng, L., Yoon, H. and Donahue, T.F. (1994)
Mol. Cell. BioI. 14,5139-5153.
95
CHAPTER 7/ PROTEIN KINASE CK2
122. Zanchin, N. L and McCarthy, I. J. (1995) J. BioI.
Chern. 270,26505-26510.
123. Bourbon, H. M., Martin-Blanco, E., Rosen, D.
and Kornberg, T. B. (1995) J. BioI. Chem. 270,
11130-11139
124. Cook, LD., Shanahan, F. and Farrel, P.J. (1994)
Virology 205, 217-227.
125. De Corte, V., Gettemans, J., De Ville, Y.,
Waelkens, E. and Vandekerckove, J. (1996)
Biochemistry 35, 5472-5480.
126. Jones, B. G., Thomas, L., Molloy, S. S., Thulin,
C. D., Fry, M. D., Walsh, K. A. and Thomas, G.
(1995) EMBO J. 14,5869-5883.
127. Norais, N., Hall, J. A., Gross, L., Tang, D.,
Kaur, S., Chamberlain, S. H., Burke, R. L. and
Marcus, F. (1996) J. Viral. 70, 5716-5719.
128. Klimzack, L. J., Collinge, M. A., Farini, D.,
Giuliano, G., Walker, J. C and Cashmore, A. R.
(1995) Plant Cell 7, 105-115.
129. Ohtsuki, K., Oh-Ishi, M., Karino, A.,
Kanekatsu, M. and Shamsa, F. (1994) Biochern.
Biophys. Res. Commun.198, 1090-1098.
130. Ohtsuki, K., Nakamura, S., Shimoyama, Y.,
Shibata, D., Munakata, H., Yoshiki, Y and
Okubo, K. (1995) J. Biochem. (Tokio) 118, 1145-
1150.
131. Cala, S.E. and Jones, L.R. (1994) J. BioI. Chern.
269,5926-5931.
132. Shen, L., Huang, K.-P., Chen, H.-C. and
Huang, F. L. (1996) Arch. Biochern. Biophys. 327,
131-141.
133. Shoshan-Barmatz, Orr, I., Meyer, H., Varsanyi,
M. and Heilmeyer L. M. G. (1996) Biochim.
Biophys. Acta 1283, 89-100.
134. Yeh, T. S., Lo, S. J., Chen, P. J. and Lee, Y. H.
(1996) J. Viral. 70,6190-6198.
135. Meggio, F., D'Agostino, D. M., Ciminale, V.,
Chieco Bianchi, L. and Pinna, L. A. (1996)
Biochern. Biophys. Res. Commun. 226,547-554.
136. Sugimoto, K. and Himeno, M. (1992) Biosci.
Biotechnol. Biochern. 56, 1174-1175.
137. Pelloux, S., Thielens, N. M., Hudry-Clergeon,
G., Petillot, Y., Filhol, O. and Arlaud, G. J.
(1996) FEBS Lett. 386,15-20.
138. Arnold, F. S., Obourn, J. D., Jaffe, H. and
Notides, A. C. (1995) J. Steroid. Biochern. BioI.
55,163-172.
139. Barroga, C. F., Stevenson, J. K., Schwatrz, E. M.
and Verma, I. M. (1995) Proc. Nat!. Acad. Sci.
USA 92, 7637-7641.
140. Chu, Z.L., McKinsey, T.A., Liu, L., Qi, X. and
Ballard, D.w. (1996) Mol. Cell. BioI. 16, 5974-
5984.
141. Das, T., Schuster, A., Schneider-Schaulies, S.
and Banerjee, A.K. (1995) Virology 211,218-226.
142. Molkentin, J. D., Li, L. and Olson, E. N. (1996)
J. BioI. Chem.271, 17199, 17204.
143. Johnson, S. E., Wang, X., Hardy, S.,
Taparowsky, E. J. and Konieczny, S. F. (1996)
Mol. Cell. BioI. 16, 1604-1613.
L. A. PINNA AND F. MEGGIO
144. Kawahire, S., Tachibana, T., Umemoto, M.,
Yoneda, Y., Imai, N., Saito, M., lchimura, T.,
Ornata, S. and Horigome, T. (1996) Exp. Cell.
Res. 222, 385-394.
145. Wong, E. V., Schaefer, A. W., Landreth, G. and
Lemmon, V. (1996) J. Neurochem. 66, 779-786.
146. Van Eynde, A., Wera, 5., Beullens, M.,
Torrekens, S., Van Leuven, F., Stalmans, W.
and Bollen, M. (1995) J. BioI. Chem. 270,28068-
28074.
147. Engel, M., Issinger, O.-G., Lascu, I., Seib, T.,
Dooley,S., Zang, K.D. and Welter, C (1994)
Biochem. Biophys. Res. Commun. 199, 1041-
1048.
148. Pai, c.Y., Chen, H.K., Sheu, H.L. and Yeh,
N.H. (1995) J. CeIl Sci. lOS, 1911-1920.
149. Vancurova, I., Paine, T. M., Lou, W. and Paine,
P. L. (1995) J. Cell Sci. lOS, 779-787.
150. Jakubowicz, T., Cytrynska, M., Kowalczyk, W.
and Gasior, E. (1993) Acta Biochim. Pol. 40, 497,
505.
151. Hashimoto, E., Takeuchi, F., Tanaka, Y. and
Yamamura, H. (1995) J. Biochem. (Tokio) 118,
453-460.
152. Pype, 5., Slegers, H., Moens, L., Merlevede, W.
and Goris, J. (1994) ]. Bioi. Chern. 269, 31457-
31465.
153. Haystead, T.A., Haystead, CM., Hu, C, Lin,
T.A. and Lawrence, J.C Jr. (1994) J. Bioi. Chern.
269,23185-23191.
154. Gallina, A. Percivalle, E., Simoncini, L.,
Revello, M. C, Gerna, G. and Milanesi, G.
(1996) J. Gen. Virol. 77, 1151-1157.
155. Zhang, Y., Beck, CA., Poletti, A., Edwards,
D.P. and Weigel, N.L. (1994) ]. Bioi. Chem.269,
31034-31040.
156. Castano, J. C, Mahillo, E., Arizti, P. and
Arribas, J. (1996) Biochemistry 35, 3782-3789.
157. Sanchez-Seco, M.P., Navarro, J., Martines, R.
and Villanueva, N. (1995) ]. Gen. Virol.76,425-
430.
158. Wu, CB., Pelech, S,L. and Veis, A. (1992) J.
Bioi. Chem. 267, 16588-16594.
159. Hockman, D. J. and Schultz, M. C (1996) Mol.
Cell. Bioi. 16,892-898.
160. Mao, C, Ray-Gallet, D., Tavitian, A., Moreau-
Gachelin, F. (1996) Oncogene 12,863-873.
161. Beretta, L., Dobransky, T. and Sobel, A. (1993)
]. BioI. Chem. 268, 20076-20084.
162. Kelliher, M.A., Seldin, D.C and Leder, P.
(1996) EMBO J. 15,5160-5166.
163. Greenwood, J.A., Scott, CW., Spreen, R.C,
Caputo, CB. and Johnson, G.V. (1994) J. Bioi.
Chem. 269, 4373-4380.
164. Kaiser, K., Stelzer, G. and Meisterernst, M.
(1995) EMBO J. 14,3520-3527.
165. Katz, D., Reginato, M. J. and Lazae, M. A.
(1995) Mol. Cell. BioI. 15,2341-2348.
166. Elizarov, S.M. and Kovaleva, G.K. (1990) Mol.
BioI. Mosk. 24, 1616-1623.
96
167. Nielander, H. B., Onofri, F., Valtorta, F.,
Schiavo, G., Montecucco, C, Greengard, P. and
Benfenati, F. (1995)]. Neurochem. 65, 1712-1720.
168. ti, M., Lyon, M. K. and Garcea, R. L. (1995) J.
BioI. Chem. 270, 26006-26011.
169. Schubert, U., Schneider, T., Kenklein, P.,
Hoffmann, K., Berthold, E., Hauser, H., Pauli,
G. and Portsmann, T. (1992) Eur. J. Biochem.
204,87~83.
170. Van Wijk, I., Burfeind, J. and Pieler, T. (1992)
Mech. Dell. 39,63-72.
171. Cairns, C and McStay, B. (1995) ,. Cell Sci.1OS,
3339-3347.
172. Meier, U. T. (1996) J. Bioi. Chern. 271, 19376-
19384.
173. Klocke, B. and Knochel, W. (1995) Mol. Cell.
Biochem.142,49-59.
174. Snell, V. and Nurse, P. (1994) EMBO J.13,
2066-2074.
175. Hartwell, L.H., Mortimer, R.K., Culotti, J. and
Culotti, M. (1973) Genetics 74, 267-286.
176. Russo, G., Vandenberg, M., Yu, I.Y., Bae, Y.-S.,
Franza, B.R. and Marshak, D.R. (1992) J. Bioi.
Chem. 267, 20317-20325.
177. Meggio, F., Boldyreff, B., Marin, 0., Issinger,
O.-G. and Pinna, L.A. (1995) Eur. J. Biochem.
230,1025-1031.
178. Mulner-Lorillon, 0., Marot, J., Cayla, X.,
Pouhle, R. and Belle, R. (1988) Eur. J. Biochem.
171,107-117.
179. Pepperkok, R., Lorenz, P., Ansorge, W. and
Pyerin, W. (1994)]. Bioi. Chem. 269,6986-6991.
180. Carrol, D. and Marshak, D.R. (1989) ]. Bioi.
Chem. 264, 7345-7348.
181. DeBenedette, M. and Snow, E.C (1991) ].
Immunol. 147,2839-2845.
182. Cardenas, M.E. and Gasser, S.M. (1993) J. Cell
Sci. 104,219-225.
183. De Paoli-Roach, A.A. (1984) J. Bioi. Chem. 259,
12144-12152.
184. Freeman, R.S. and Donoghue, D.J. (1991)
Biochemistry 30, 2293-.2302
185. Brautigan, T.L., Sunwoo, I., Labbe, J.C,
Fernandez, A. and Lamb, N.J. (1990) Nature
344,74-78.
186. Schneider, H.R., Reichert, G. and Issinger, 0.-
G. (1986) Eur. J. Biochem. 161, 733-738.
187. Perez, M., Grande, J. and Itarte, E. (1987) J.
Biochem. 170, 493-498.
188. Durand, P., Vilgrain, J., Chambaz, E.M. and
Saez, J.M. (1987) Mol. Cell. Endocrinol. 53, 195-
202.
189. Girault, A.-J., Hemmings, H.C, Jr., Zorn, 5tH.,
Gustafson, E.L. and Greengard, P. (1990) J.
Neurochem.55,l772-1783.
190. Maridor, G., Park, W., Krek, W. and Nigg, E.A.
(1991) J. Bioi. Chem. 266, 2362-2368.
191. Hu, E. and Rubin, CS. (1990) J. Bioi. Chem. 265,
5072-5080.

192. Mestres, P., Boldyreff, B., Ebensperger, c.,
Hameister, H. and Issinger, O.-G. (1994) Acta
anatomica 149, 13-.20.
193. Perez, M., Grande, J. and Itarte, E. (1988) FEBS
Lett. 238, 273-276.
194. Pancetti, F., Bosser, R., Harte, E. and Bachs, O.
(1996) Biochem. Biophys. Res. Commun. 218, 35-
39.
195. Lamprecht, S.A., Schwartz, B., Avigdor, A. and
Guberman, R. (1990) Anticancer Res. 10, 773-
778.
196. Miinstermann, U., Fritz, G., Seitz, G., Yiping,
L., Schneider, H.R. and Issinger, O.-G. (1990)
Eur. J. Biochem.189, 251-257.
197. Schneider, H.R. and Issinger, O.-G. (1989)
Biotec. Europe 6, 82-88.
198. Pistorius, K., Seitz, G., Remberger, K. and
Issinger, O.-G. (1991) Onkologie 14, 256-260.
199. Boldyreff, B., Lu, Y. and Issinger, O.-G. (1991)
Onkologie Aktue1l8, 124-129.
97
CHAPTER 7/ PROTEIN KINASE CK2
200. Issinger, O.-G. and Boldyreff, B. (1992) In
Recent Advances in Cellular and Molecular
Biology (Wegmann, R. and Wegmann, M. eds.)
vol. 4, 17-22, Peters Press, Leuven, Belgium.
201. Venice, 5., Davis, A.T., Goueli, S.A., Akdas, A.,
Limas, C. and Ahmed, K. (1994) The Prostate
24,11-16.
202. Pena, J.M., ltarte, E., Domingo, A. and Cusso,
R. (1983) Cancer Res. 43, 1172-1175.
203. Brunati, A.M., Saggioro, D., Chieco Bianchi, L.
and Pinna, L.A. (1986) FEBS Lett. 206,59-63.
204. ole-MoiYoi, O.K., Brown, W.c., lams, K.P.,
Nayar, A., Tsukamoto, T. and Mackin, M.D.
(1993) EMBO J. 12, 1621-1631.
205. Kelliher, M.A., Seldin D.C. and Leder, P (1996)
EMBO J. 15, 5160-5166.
206. Taylor, 5.5. and Radzio-Andzelm, E. (1994)
Structure, 2,345-355.