See

discussions, stats, and author profiles for this publication at: https://www.researchgate.net/publication/40766874

From signaling pathways to microtubule

dynamics: The key players

Article in Current opinion in cell biology · February 2010

DOI: 10.1016/j.ceb.2009.11.008 · Source: PubMed

CITATIONS READS

93 170

1 author:

Sandrine Etienne-Manneville
Institut Pasteur International Network
64 PUBLICATIONS 8,368 CITATIONS

SEE PROFILE

All content following this page was uploaded by Sandrine Etienne-Manneville on 22 February 2017.

The user has requested enhancement of the downloaded file. All in-text references underlined in blue are added to the original document
and are linked to publications on ResearchGate, letting you access and read them immediately.
 Available online at www.sciencedirect.com
From signaling pathways to microtubule dynamics: the key
players
Sandrine Etienne-Manneville
Microtubules are highly dynamic structures whose regulation is
crucial for cell division, cell polarity, cell migration, or neuronal
differentiation. Because they contribute to most cellular
functions, they must be regulated in response to extracellular
and intracellular signals. The parameters of microtubule
dynamics are numerous and complex and the connection
between signaling pathways and regulation of microtubule
dynamics remain obscure. Recent observations reveal key
players that can both integrate the diversity of signaling
cascades and directly influence microtubule dynamics. I review
here how modifications of the tubulin dimer, tubulin modifying
enzymes, and microtubule-associated proteins are directly
involved in the regulation of microtubule behavior and
functions.
Address
Institut Pasteur, Cell Polarity and Migration Group and CNRS URA 2582,
25 rue du Dr Roux, 75724 Paris Cedex 15, France
Corresponding author: Etienne-Manneville, Sandrine
(sandrine.etienne-manneville@pasteur.fr)
Current Opinion in Cell Biology 2010, 22:104–111
This review comes from a themed issue on
Cell structure and dynamics
Edited by Arshad Desai and Marileen Dogterom
Available online 22nd December 2009
0955-0674/$ – see front matter
# 2009 Elsevier Ltd. All rights reserved.
DOI 10.1016/j.ceb.2009.11.008
Introduction
Microtubules are stiff, cylindrical polymers of a-tubulins
and b-tubulins. Head-to-tail association of a–b hetero-
dimers forms linear protofilaments. Tubulin sheets of 13
protofilaments are sealed by lateral interactions between
the most external protofilaments to form the fully poly-
merized, hollowed microtubules. The polarized arrange-
ment of the tubulin dimers gives the polymer a
molecular polarity with a ‘minus’ (a-tubulin) end and
a ‘plus’ (b-tubulin) end. Microtubules form the stable
rigid core of complex structures such as axonemes in cilia
and flagella. But, like microfilaments, microtubules can
undergo rapid polymerization and depolymerization. In
interphase animal cells, the microtubule network gener-
ally extends as a radial array from the microtubule
organizing center, where most minus-ends are anchored
Current Opinion in Cell Biology 2010, 22:104–111
and which often corresponds to the centrosome, to the
periphery of the cell where microtubule plus-ends con-
stantly explore the cytoplasm. The microtubule plus-
ends display what is called dynamic instability, a funda-
mental process which encompasses a succession of slow
polymerization and rapid depolymerization phases sep-
arated by transitions. The transition between polymer-
ization and shortening is called a catastrophe and the
transition between depolymerization and growth is
called a rescue. Dynamic instability allows microtubules
to search and find the various elements of the cell
architecture. Microtubules serve as tracks for dynein
and kinesin molecular motors that deliver membrane
vesicles, constituent proteins, and regulatory factors. In
addition to their fundamental role in cell division, micro-
tubules also participate in cell shape changes and
directed motility in migrating cells or neuronal growth
cones [1]. During the last decade, it has become clear
that the microtubule network, like the actin cytoskele-
ton, is regulated by a multitude of stimuli. A number of
extracellular or intracellular signals including stimu-
lation by soluble factors, cellular interactions, and
physical constraints are shown to control microtubule
organization [2–5]. Recent and comprehensive reviews
of these pathways can be found in [1,6–9]. Formation and
positioning of the mitotic spindle as well as microtubule-
dependent neuronal differentiation and migration have
brought forward new signaling pathways regulating
microtubule dynamics and functions. Polarity proteins,
such as MARK/Par1 and the Wnt target GSK3 kinase, or
Aurora B which controls spindle pole assembly, and more
general signaling components, such as heterotrimeric G
proteins, modulate the activity of microtubule-associ-
ated proteins (MAPs) that act at the interface between
signaling cascades and microtubules. I will focus here on
the recent observations that point to essential molecular
targets responsible for the coupling between signaling
pathways and microtubule dynamics.
Tubulin, more than a solid brick
The most direct way to affect microtubule dynamics is to
control the accessibility or the conformation of its build-
ing brick the a–b-tubulin dimer. Tubulin dimers are
formed by an a-tubulin constitutively associated to a
guanosine triphosphate (GTP) and a b-tubulin which
cycles between a GTP-bound form and a guanosine
diphosphate (GDP)-bound form (Figure 1a). Polymeriz-
ation occurs by the addition of tubulin dimers and also
oligomers that are formed prior their incorporation in the
growing microtubules [10].
www.sciencedirect.com
 Regulation of microtubule dynamics Etienne-Manneville 105

Figure 1

Protofilament conformation is a key parameter in microtubule dynamics. (a) Microtubule polymerization occurs by head-to-tail addition of a–b tubulin
heterodimers. Binding of GTP to b-tubulin and then integration to the protofilament straighten the dimers. (b) Microtubule polymerization. The plus-end
displays a sheet-like conformation which is eventually sealed to form the microtubule. The straight conformation of the growing protofilaments (formed
by polymerization of GTP-bound dimers, green b-tubulin) protects the microtubule from depolymerization. Following polymerization, GTP hydrolysis
occurs (red b-tubulin), but some GTP-bound b-tubulins can remain along the microtubule (GTP-remnant). (c) During depolymerization, protofilaments
containing GDP–b-tubulin are curved outwards. Depolymerization can stop where GTP-remnants form a stable ring of straight dimers (lower panel).
Some of the molecules (MAPs, XMAP215, and MCAK) binding to microtubules or tubulin dimers to control their conformation are represented.

A critical parameter in microtubule dynamics is the
conformation of the tubulin protofilaments (Figure 1).
Association of a GTP or a GDP with the b-tubulin greatly
affects the conformation of the tubulin dimer and controls
microtubule assembly and dynamics [11]. When b-tubu-
lin is bound to GTP, the corresponding dimer displays a
straighter conformation which facilitates the extension of
the protofilaments [11,12]. Following association with the
microtubule wall, the tubulin dimer further straightens
[13] (Figure 1a). The plus-ends of growing microtubules
are therefore formed of GTP-bound dimers included in
straight protofilaments which organize as a sheet before
forming the hollow tube [14] (Figure 1b). After polymer-
ization, hydrolysis of the GTP occurs with time and
usually when the dimer is included within the microtu-
bule. Although the phosphate is not released, the GDP-
Pi-bound dimer exhibits an increased curvature, making
the protofilaments more prone to depolymerization. In
the absence of GTP-bound tubulin at the end of the
microtubule, the protofilaments curve outwards losing
lateral contacts and depolymerizing rapidly (Figure 1c).
When the microtubule is growing, the delayed hydrolysis
of the GTP results in the existence of a ‘GTP-cap’ that
is thought to stabilize the open-sheet conformation of

www.sciencedirect.com Current Opinion in Cell Biology 2010, 22:104–111

106 Cell structure and dynamics
growing microtubule plus-ends and to prevent microtu-
bule shrinkage and catastrophe [15,16]. The size of the
GTP-cap depends on the polymerization rate and may
also vary with the GTPase activity of tubulin. Using a
recombinant antibody specifically recognizing GTP–
tubulin, Dimitrov et al. have been able to localize
GTP–tubulin in vivo [17]. In addition to the expected
GTP-cap, GTP-bound tubulin is also present along the
microtubules (Figure 1b) sometimes forming long
stretches that seem to correlate with microtubule bund-
ling. GTP–tubulin also appears as discrete dot-like
regions. These GTP–tubulin clusters seem to, at least
temporarily, block microtubule shrinking as they corre-
spond to the location of rescue events (Figure 1c). GTP-
remnants form during in vitro polymerization suggesting
that their presence does not require any other external
factors. The regulation of the tubulin GTPase activity is
likely to be critical in the control of microtubule dynamics
and stability.
Several proteins regulate microtubule dynamics by mod-
ifying the accessibility and the conformation of tubulin
dimers. The microtubule destabilizing drug colchicine
prevents curved tubulin from adopting a straight structure
and inhibits assembly [18]. Stathmin inhibits microtubule
growth by sequestering soluble tubulin and decreasing
the concentration of accessible tubulins in proximity of
microtubule-ends. Stathmin binds with tubulin hetero-
dimers in a curved complex which prevents incorporation
of the complexed tubulins into microtubules. Stathmin
association with tubulin is inhibited by phosphorylation.
Among the four phosphorylable serine residues, Ser16
and Ser63 seem to be the most critical [19]. Regulation of
Stathmin by phosphorylation on Ser16 by the Rac effector
PAK1 has been involved in the regulation of the micro-
tubule network in migrating cells and developing neurons
[20,21]. Phosphorylation of the same serine residue by
Aurora B has also been involved during mitotic spindle
assembly [22]. Other kinases like Calmodulin-dependent
kinase, Erk MAP kinase, or PKA have also been shown to
phosphorylate Stathmin, which appears as a key target for
a large variety of signaling pathways. The microtubule
destabilizer MCAK stabilizes the curved conformation of
the protofilaments at the microtubule-end and thereby
favors depolymerization [23] (Figure 1c). During mitosis,
phosphorylation by Aurora B leads to MCAK recruitment
to spindle poles and promotes bipolar spindle assembly
[24].
In contrast to microtubule destabilizers, XMAP215 inter-
acts with tubulin dimers to facilitate their addition to
microtubule plus-ends and promote microtubule growth
[25,26]. In the same way as taxol stabilizes microtubules
by straightening the GDP–tubulin protofilaments [12],
MAPs stabilize microtubule by interacting with consecu-
tive dimers in the microtubule lattice (Figures 1b and 2a).
Once again, phosphorylation appears as a means to
Current Opinion in Cell Biology 2010, 22:104–111
regulate microtubule interaction and thus the function
of these proteins. Kinases of the Par1/MARK family
phosphorylate the Tau protein and related MAPs on a
conserved KXGS motif to induce their dissociation from
microtubules. This regulatory mechanism is extensively
studied as it participates to cell polarity in the Caenor-
habditis elegans embryo and in epithelial cells, and also to
neuronal differentiation and neurodegenerative diseases
such as Alzheimer’s disease (see for a review [7]). The
GSK3 kinase, a crucial intermediate in Wnt signaling and
in cell polarity, is also involved in this regulatory mechan-
ism not only by phosphorylating Tau or MAPs on residues
different from those regulated by MARK but also by
modulating MARK activity by phosphorylation [27].
The precise pattern of phosphorylation achieved by the
convergence of multiple pathways on microtubule-associ-
ated proteins may allow a fine tuning of microtubule
stability. For the moment, only very few reports indicate
a possible regulation of the tubulin GTPase activity. Inter-
estingly, the alpha subunit of heterotrimeric G proteins
activates the GTP hydrolysis [28]. It may thus control
microtubule polymerization at the plus-ends and also
influence the stability of the overall microtubule by con-
trolling the position of GTP-remnants. Since heterotri-
meric G proteins play a key role in microtubule-dependent
asymmetric positioning of the mitotic spindle in C. elegans
and Drosophila [29], it is tempting to hypothesize that
regulation of tubulin GTPase activity is at the interface
between polarity signaling pathways and the regulation of
microtubule dynamics and anchoring at the cell cortex.
Tubulin post-translational modifications, road
signs along the lattice
Tubulins can be modified by several post-translational
modifications, including acetylation, detyrosination, and
deglutamylation of detyrosinated tubulin (D2 modifi-
cation), poly-glutamylation and poly-glycylation [30]
(Figure 2). These modifications occur when tubulin is
included into a microtubule. Except D2 modification,
these reactions are reversible.
Detyrosination is catalyzed, once tubulin is incorporated
into the microtubule lattice, by cytosolic carboxypepti-
dases which are not yet fully characterized [30]. Tubulin
tyrosine ligase (TTL) catalyzes the reverse reaction on
soluble tubulin only. Detyrosination was initially thought
not to affect the dynamics of microtubules [31] but rather
to mark stable microtubules and affect microtubule inter-
action with molecular motors and in particular with kine-
sins [32,33] (Figure 2a). Depletion of TTL has important
consequences on neuron differentiation while the
microtubule network appears globally unaffected [34].
However, it now seems that the accumulation of detyr-
osinated tubulin alters the binding of EB1 and Clip170,
which are key regulators of microtubule dynamics [35,36],
suggesting that tyrosination can at least indirectly affect
microtubule dynamics.
www.sciencedirect.com
 Regulation of microtubule dynamics Etienne-Manneville 107

Figure 2

Tubulin structure and post-translational modifications control microtubule interactions with partner proteins. (a) Along a polymerized microtubule,
post-translational modifications such as detyrosination (carboxy-terminal EE, light blue), (poly)glutamylation (turquoise EEE), and (poly)glycylation
(mainly in axonemes, purple GGG), regulate microtubule association with motors (kinesins) or MAPs (microtubule-associated proteins). Acetylation
that occurs on the inner surface of the microtubule is not shown. (b) EB1 (bright green) recognizes the conformation of the growing microtubule plus-
end. For simplification, EB1 dimers are not depicted. More microtubule plus-end tracking proteins (+TIPs) are recruited to the plus-ends by binding
directly to tyrosinated a-tubulin (carboxy-terminal EEY) or to EB1 sequences.

Acetylation, like detyrosination, was initially thought to
simply mark stable, long-lived microtubules. In agree-
ment, the tubulin deacetylase HDAC6-deficient mice
show hyperacetylated tubulin in most tissues but develop
normally [37]. However, evidence is now accumulating
that tubulin acetylation may play a role in microtubule
functions [38,39]. Creppe et al. have recently uncovered a
putative tubulin acetylase activity associated with the
multisubunit histone acetyltransferase elongator complex
which is involved in dendrite and axonal growth [40].
This may be due to the fact that such tubulin modifying
enzymes have multiple substrates in addition to tubulin
[41,42]. However, in this case, expression of a nonacety-
latable a-tubulin mimics the effects of acetylase
depletion supporting the direct role of tubulin acetylation
in migration and branching of cortical neurons. In fact,
although acetylation occurs on K40, an amino acid
exposed in the microtubule lumen, it has been shown
to affect the interactions of molecular motors with the
microtubule exterior wall [32,33].
The impact of other tubulin modifications on microtubule
dynamics has not yet been clearly determined. The
recent identification of a large family of TTL-Like
(TTLL) genes encoding poly-glutamylases (TTLL 1,
4–7, 11, and 13) and poly-glycylases (TTLL 3, 8, 10, and
12) [43,44,45] has given the opportunity to determine
the effects of such modifications on microtubule
dynamics. These TTLLs differ in their specificity for
a-tubulin or b-tubulin and in catalyzing the initiation or
the elongation of the peptidic side chain [43]. The
accumulation of these post-translational modifications

www.sciencedirect.com Current Opinion in Cell Biology 2010, 22:104–111

108 Cell structure and dynamics
in microtubules of cilia, flagella and mitotic spindle is now
echoed by the fact that TTLLs appear to be required for
the organization and function of microtubules in cilia and
flagella [46,47]. Like detyrosination, poly-glutamylation,
and poly-glycylation occur on the carboxy-terminal region
of tubulins that are likely to influence the binding of
microtubule-interacting proteins. Association of kinesin
motors with microtubules is affected by these modifi-
cations (Figure 2a). Kinesin-1 requires b-tubulin poly-
modifications [32], whereas kinesin-3 is sensitive to a-
tubulin poly-glutamylation [48]. In addition severing
proteins that can dramatically affect microtubule
dynamics are also influenced by b-tubulin poly-modifi-
cations [49,50].
Accumulation of tubulin modifications in neuronal cells
[51], altered expression levels of TTL during tumorigen-
esis together with the existence of new genetic tools will
probably encourage many studies to help us understand
how the enzymes responsible for these modifications are
regulated and further determine the consequences of
these microtubule modifications. Although how tubulin
post-translational modifications can directly affect micro-
tubule dynamics remains to be determined, it is clear that
they can affect microtubule interactions with their regu-
latory partners and thus indirectly regulate microtubule
polymerization. A remaining and important question here
is how the activity and the localization of the tubulin
modifying enzymes are regulated.
EB1, master at the plus-ends
Proteins specifically accumulating at the plus-ends, called
+TIPs, such as end-binding (EB) proteins, adenomatous
polyposis coli (APC), or Clip170 are fundamental regu-
lators of microtubule plus-end dynamics and have there-
fore been the center of attention [52] (please also see the
review by K Slep in this issue). The EB proteins EB1 and
EB3 have recently appeared as key proteins of the plus-
ends. In contrast to most +TIPs, EBs can track micro-
tubule plus-ends in an autonomous manner [53]. Like its
yeast ortholog Mal3p, EB1 interacts preferentially with
the free lateral sides of protofilaments and therefore
shows an enhanced affinity for the plus-end structure
of a polymerizing microtubule as opposed to the older
closed microtubule lattice [53,54] (Figure 2b). EB1
directly stimulates microtubule nucleation and growth
by facilitating the incorporation of tubulin dimers and
promotes microtubule sheet closure by strengthening
lateral interactions between straight adjacent protofila-
ments [55,56]. On the one hand, the accelerated closure
of the microtubule sheet results in an increase in cata-
strophe events, probably because the conformation of the
unsealed microtubule-ends stabilizes the plus-ends. On
the other hand, high concentration of EB1 increases the
rescue frequency by interacting with and stabilizing the
microtubule wall [55]. In vivo, EB1 rather acts as an
inhibitor of catastrophes suggesting that the concen-
Current Opinion in Cell Biology 2010, 22:104–111
tration or activity of the cytosolic EB1 pool may be locally
controlled [57]. The extent of the GTP-cap in parallel to
the regulation of the protofilament curvature at plus-ends
is likely to control EB1 recruitment.
In addition to its direct role in the regulation of micro-
tubule dynamics, EB1 acts as a loading factor of other
+TIPs. The wide variety of EB1 partners can fall into two
distinct categories: proteins containing a cytoskeleton-
associated protein-glycine-rich (CAP-Gly) domain, like
Clip170 and p150Glued [56] and proteins including an
EB1-binding domain composed of a short SxIP motif
([58] and K Slep, this issue). Clip170, initially thought
to autonomously track microtubule plus-ends, in fact
recognizes the EB1–tubulin complex present at the
plus-ends [59] and p150Glued may function similarly.
The positively charged CAP-Gly domain interacts with
the EEY/F carboxy-terminal acidic motif which is present
both on EB1 and tyrosinated tubulins (Figure 2b) [60].
Since detyrosination of tubulin occurs after polymeriz-
ation, tyrosinated tubulin is preferentially found at the
plus-ends. The accumulation of Clip170 and p150Glued at
the plus-ends of microtubules is likely to result from their
direct binding to tyrosinated tubulin and from their
association to plus-end tracking EB1. In TTL-null neurons
which do not contain tyrosinated tubulin, Clip170 is mis-
localized whereas EB1 localization is unaffected [34],
suggesting that tyrosinated tubulin remains the primary
signal for CAP-Gly protein tracking properties and that
regulation of tubulin carboxypeptidase may also contribute
to control dynamics at microtubule plus-ends.
Proteins such as APC, CLASPs, or MCAK, which were all
known to bind EB1 but did not show any obvious
sequence similarities, all present a SxIP motif [58,61].
CDK5RAP2 has also been shown to interact via the same
interaction motif [62]. The carboxy-terminal domain of
EB1 appears as a mini docking platform as the list of
interacting partners, participating in microtubule plus-
end function, lengthens (Figure 2b). Different +TIPs
may be recruited simultaneously on EB1 dimers (K Slep,
this issue). The EEY/F binding site of the CAP-Gly
proteins [63] is separated by only eight residues from
the site recognizing the SxIP motif suggesting that the
two categories of proteins may interact and either com-
pete or cooperate to control microtubule plus-end
dynamics. Regulation of interactions at the plus-ends
can affect both microtubule dynamics and microtubule
functions. Can signaling pathways control the nature and
the quantity of +TIPs present on a microtubule extre-
mity? The regulation is likely to be very local as micro-
tubules not only have different functions in different
regions of the cells but can also display various properties
in a same cell area. Phosphorylation near the EB1-binding
domains of +TIPs also regulates their interaction with
EB1 [58,64]. Phosphorylation also affects the inter-
action between MCAK and EB1-TIP150 preventing
www.sciencedirect.com

MCAK recruitment to microtubule plus-ends [65]. Some
+TIPs can also be found along microtubules, where they
may facilitate rescue events. Whether GTP-remnants
along the microtubule exhibit a peculiar structure that
resembles a plus-end and facilitates EB1 binding is still
unclear. It is nevertheless tempting to consider that
localization of +TIPs along microtubules may result from
a local inhibition of tubulin GTPase activity which would
result in a higher probability of rescue [17,56].
Conclusions
To fulfill their cellular functions, microtubules must
respond to their environment. Recent observations
demonstrate that microtubules are subjected to re-
arrangements that follow changes in cell shape or intra-
cellular architecture [2–4]. The coupling between
extracellular or intracellular signals and the regulation
of microtubule dynamics and functions must be finely
tuned. This tuning relies on essential players which are
first, the accessibility and the conformational state of
tubulin dimers; second, the tubulin modifier enzymes
which control tubulin interactions with microtubule
motors or regulators; and third, the +TIPs which assemble
around EB1. These critical microtubule regulators are the
targets of signaling pathways that act via kinases such as
MARKs or GSK3, heterotrimeric G proteins and other yet
unidentified molecules.
By controlling the conformation of protofilaments and
protein interactions along and at the ends of microtu-
bules, signaling pathways regulate microtubule dynamics
and also functions. The structure of depolymerizing
microtubules is recognized by fibrils that can use micro-
tubule shortening to drive chromosome motion toward
the spindle poles [66]. Modulation of kinesin association
to microtubules by tubulin post-translational modifi-
cations allows the delivery of molecules, complexes or
membrane vesicles to defined cell regions. +TIPs are also
essential to mediate the interaction between microtu-
bules and various subcellular structures such as orga-
nelles, actin cytoskeleton, and the cell cortex and can
help to locally concentrate regulatory molecules. These
interactions, which can involve another layer of mol-
ecules, are essential in coordinating microtubule
dynamics with actin cytoskeleton rearrangements [3,67]
or remodeling of cell interactions with the extracellular
matrix [68]. More recently, interaction between micro-
tubule-ends and cell–cell junctions have been shown to
promote a bidirectional signaling allowing the regulation
of microtubule dynamics [69,70] and the formation or
stabilization of cellular junctions [71–73]. The key players
that couple microtubule dynamics and signaling pathways
seem to have been identified. Further characterization
and better understanding of their regulation is now
needed to clarify the role of microtubules in cellular
functions and dysfunctions, developmental processes
and diseases.
www.sciencedirect.com
Regulation of microtubule dynamics Etienne-Manneville 109
Acknowledgements
SEM is supported by the Centre National de la Recherche Scientifique, the
Institut Pasteur, the Fondation de France and La Ligue contre le Cancer
and is a member of the EMBO YIP.
References and recommended reading
Papers of particular interest, published within the period of review,
have been highlighted as:
 of special interest
 of outstanding interest
1. Lansbergen G, Akhmanova A: Microtubule plus end: a hub of
cellular activities. Traffic 2006, 7:499-507.
2. Minc N, Bratman SV, Basu R, Chang F: Establishing new sites of
polarization by microtubules. Curr Biol 2009, 19:83-94.
3. Terenna CR, Makushok T, Velve-Casquillas G, Baigl D, Chen Y,
Bornens M, Paoletti A, Piel M, Tran PT: Physical mechanisms
redirecting cell polarity and cell shape in fission yeast. Curr Biol
2008, 18:1748-1753.
4. Dumont S, Mitchison TJ: Compression regulates mitotic spindle
length by a mechanochemical switch at the poles. Curr Biol
2009, 19:1086-1095.
5. Athale CA, Dinarina A, Mora-Coral M, Pugieux C, Nedelec F,
Karsenti E: Regulation of microtubule dynamics by reaction
cascades around chromosomes. Science 2008, 322:1243-1247.
6. Barth AI, Caro-Gonzalez HY, Nelson WJ: Role of adenomatous
polyposis coli (APC) and microtubules in directional cell
migration and neuronal polarization. Semin Cell Dev Biol 2008,
19:245-251.
7. Matenia D, Mandelkow EM: The tau of MARK: a polarized view
of the cytoskeleton. Trends Biochem Sci 2009, 34:332-342.
8. Roychowdhury S, Rasenick MM: Submembraneous
microtubule cytoskeleton: regulation of microtubule assembly
by heterotrimeric G proteins. FEBS J 2008, 275:4654-4663.
9. Akhmanova A, Stehbens SJ, Yap AS: Touch, grasp, deliver and
control: functional cross-talk between microtubules and cell
adhesions. Traffic 2009, 10:268-274.
10. Mozziconacci J, Sandblad L, Wachsmuth M, Brunner D,
Karsenti E: Tubulin dimers oligomerize before their
incorporation into microtubules. PLoS ONE 2008, 3:e3821.
11. Wang HW, Nogales E: Nucleotide-dependent bending flexibility
of tubulin regulates microtubule assembly. Nature 2005,
435:911-915.
12. Elie-Caille C, Severin F, Helenius J, Howard J, Muller DJ,
Hyman AA: Straight GDP–tubulin protofilaments form in the
presence of taxol. Curr Biol 2007, 17:1765-1770.
13. Rice LM, Montabana EA, Agard DA: The lattice as allosteric
 effector: structural studies of alphabeta- and gamma-tubulin
clarify the role of GTP in microtubule assembly. Proc Natl Acad
Sci U S A 2008, 105:5378-5383.
Based on gamma-tubulin structure analysis, this report shows that
incorporation of a tubulin dimer in a protofilament is responsible for most
of the conformational change in the dimer and that GTP loading has a less
important effect.
14. Gardner MK, Hunt AJ, Goodson HV, Odde DJ: Microtubule
assembly dynamics: new insights at the nanoscale. Curr Opin
Cell Biol 2008, 20:64-70.
15. Chretien D, Jainosi I, Taveau JC, Flyvbjerg H: Microtubule’s
conformational cap. Cell Struct Funct 1999, 24:299-303.
16. Janosi IM, Chretien D, Flyvbjerg H: Structural microtubule cap:
stability, catastrophe, rescue, and third state. Biophys J 2002,
83:1317-1330.
17. Dimitrov A, Quesnoit M, Moutel S, Cantaloube I, Pous C, Perez F:
Detection of GTP–tubulin conformation in vivo reveals a role
for GTP remnants in microtubule rescues. Science 2008,
322:1353-1356.
Current Opinion in Cell Biology 2010, 22:104–111
110 Cell structure and dynamics
18. Ravelli RB, Gigant B, Curmi PA, Jourdain I, Lachkar S, Sobel A,
Knossow M: Insight into tubulin regulation from a complex with
colchicine and a stathmin-like domain. Nature 2004,
428:198-202.
19. Manna T, Thrower DA, Honnappa S, Steinmetz MO, Wilson L:
Regulation of microtubule dynamic instability in vitro by
differentially phosphorylated stathmin. J Biol Chem 2009,
284:15640-15649.
20. Watabe-Uchida M, John KA, Janas JA, Newey SE, Van Aelst L:
The Rac activator DOCK7 regulates neuronal polarity through
local phosphorylation of stathmin/Op18. Neuron 2006,
51:727-739.
21. Wittmann T, Bokoch GM, Waterman-Storer CM: Regulation of
microtubule destabilizing activity of Op18/stathmin
downstream of Rac1. J Biol Chem 2004, 279:6196-6203.
22. Gadea BB, Ruderman JV: Aurora B is required for mitotic
chromatin-induced phosphorylation of Op18/Stathmin. Proc
Natl Acad Sci U S A 2006, 103:4493-4498.
23. Hertzer KM, Walczak CE: The C-termini of tubulin and the
 specific geometry of tubulin substrates influence the
depolymerization activity of MCAK. Cell Cycle 2008,
7:2727-2737.
MCAK binds to the plus-ends of microtubules and stabilizes a curved
conformation of the protofilaments to help their disassembly.
24. Zhang X, Ems-McClung SC, Walczak CE: Aurora A
phosphorylates MCAK to control ran-dependent spindle
bipolarity. Mol Biol Cell 2008, 19:2752-2765.
25. Brouhard GJ, Stear JH, Noetzel TL, Al-Bassam J, Kinoshita K,
Harrison SC, Howard J, Hyman AA: XMAP215 is a processive
microtubule polymerase. Cell 2008, 132:79-88.
26. Slep KC, Vale RD: Structural basis of microtubule plus end
tracking by XMAP215, CLIP-170, and EB1. Mol Cell 2007,
27:976-991.
27. Timm T, Balusamy K, Li X, Biernat J, Mandelkow E,
Mandelkow EM: Glycogen synthase kinase (GSK) 3beta directly
phosphorylates Serine 212 in the regulatory loop and inhibits
microtubule affinity-regulating kinase (MARK) 2. J Biol Chem
2008, 283:18873-18882.
28. Roychowdhury S, Panda D, Wilson L, Rasenick MM: G protein
alpha subunits activate tubulin GTPase and modulate
microtubule polymerization dynamics. J Biol Chem 1999,
274:13485-13490.
29. Gonczy P: Mechanisms of asymmetric cell division: flies
and worms pave the way. Nat Rev Mol Cell Biol 2008,
9:355-366.
30. Hammond JW, Cai D, Verhey KJ: Tubulin modifications and their
cellular functions. Curr Opin Cell Biol 2008, 20:71-76.
31. Palazzo A, Ackerman B, Gundersen GG: Cell biology: tubulin
acetylation and cell motility. Nature 2003, 421:230.
32. Reed NA, Cai D, Blasius TL, Jih GT, Meyhofer E, Gaertig J,
Verhey KJ: Microtubule acetylation promotes kinesin-1
binding and transport. Curr Biol 2006, 16:2166-2172.
33. Bulinski JC: Microtubule modification: acetylation speeds
anterograde traffic flow. Curr Biol 2007, 17:R18-20.
34. Erck C, Peris L, Andrieux A, Meissirel C, Gruber AD, Vernet M,
Schweitzer A, Saoudi Y, Pointu H, Bosc C et al.: A vital role of
tubulin-tyrosine-ligase for neuronal organization. Proc Natl
Acad Sci U S A 2005, 102:7853-7858.
35. Peris L, Thery M, Faure J, Saoudi Y, Lafanechere L, Chilton JK,
Gordon-Weeks P, Galjart N, Bornens M, Wordeman L et al.:
Tubulin tyrosination is a major factor affecting the recruitment
of CAP-Gly proteins at microtubule plus ends. J Cell Biol 2006,
174:839-849.
36. Badin-Larcon AC, Boscheron C, Soleilhac JM, Piel M, Mann C,
Denarier E, Fourest-Lieuvin A, Lafanechere L, Bornens M, Job D:
Suppression of nuclear oscillations in Saccharomyces
cerevisiae expressing Glu tubulin. Proc Natl Acad Sci U S A
2004, 101:5577-5582.
Current Opinion in Cell Biology 2010, 22:104–111
37. Zhang Y, Kwon S, Yamaguchi T, Cubizolles F, Rousseaux S,
Kneissel M, Cao C, Li N, Cheng HL, Chua K et al.: Mice lacking
histone deacetylase 6 have hyperacetylated tubulin but are
viable and develop normally. Mol Cell Biol 2008, 28:1688-1701.
38. Matsuyama A, Shimazu T, Sumida Y, Saito A, Yoshimatsu Y,
Seigneurin-Berny D, Osada H, Komatsu Y, Nishino N, Khochbin S
et al.: In vivo destabilization of dynamic microtubules by
HDAC6-mediated deacetylation. EMBO J 2002, 21:6820-6831.
39. Tran AD, Marmo TP, Salam AA, Che S, Finkelstein E, Kabarriti R,
Xenias HS, Mazitschek R, Hubbert C, Kawaguchi Y et al.: HDAC6
deacetylation of tubulin modulates dynamics of cellular
adhesions. J Cell Sci 2007, 120:1469-1479.
40. Creppe C, Malinouskaya L, Volvert ML, Gillard M, Close P,
 Malaise O, Laguesse S, Cornez I, Rahmouni S, Ormenese S et al.:
Elongator controls the migration and differentiation of cortical
neurons through acetylation of alpha-tubulin. Cell 2009,
136:551-564.
This report shows the role of the multisubunit histone acetyltransferase
elongator complex (Elp1–Elp6) in microtubule acetylation in differentiating
projection neurons. The catalytic Elp3 subunit promotes a-tubulin acet-
ylation. The authors also show that inhibition of tubulin acetylation by
expression of a nonacetylable mutant alters neuronal branching and
migration.
41. Kovacs JJ, Murphy PJ, Gaillard S, Zhao X, Wu JT, Nicchitta CV,
Yoshida M, Toft DO, Pratt WB, Yao TP: HDAC6 regulates Hsp90
acetylation and chaperone-dependent activation of
glucocorticoid receptor. Mol Cell 2005, 18:601-607.
42. Zhang X, Yuan Z, Zhang Y, Yong S, Salas-Burgos A, Koomen J,
Olashaw N, Parsons JT, Yang XJ, Dent SR et al.: HDAC6
modulates cell motility by altering the acetylation level of
cortactin. Mol Cell 2007, 27:197-213.
43. van Dijk J, Rogowski K, Miro J, Lacroix B, Edde B, Janke C: A
 targeted multienzyme mechanism for selective microtubule
polyglutamylation. Mol Cell 2007, 26:437-448.
The manuscript reports the identification of six mammalian polyglutamy-
lases which differ by the exact type of tubulin modifications they catalyze.
44. Ikegami K, Mukai M, Tsuchida J, Heier RL, Macgregor GR,
Setou M: TTLL7 is a mammalian beta-tubulin polyglutamylase
required for growth of MAP2-positive neurites. J Biol Chem
2006, 281:30707-30716.
45. Janke C, Rogowski K, Wloga D, Regnard C, Kajava AV, Strub JM,
Temurak N, van Dijk J, Boucher D, van Dorsselaer A et al.: Tubulin
polyglutamylase enzymes are members of the TTL domain
protein family. Science 2005, 308:1758-1762.
46. Pathak N, Obara T, Mangos S, Liu Y, Drummond IA: The zebrafish
fleer gene encodes an essential regulator of cilia tubulin
polyglutamylation. Mol Biol Cell 2007, 18:4353-4364.
47. Wloga D, Webster DM, Rogowski K, Bre MH, Levilliers N, Jerka-
Dziadosz M, Janke C, Dougan ST, Gaertig J: TTLL3 Is a tubulin
glycine ligase that regulates the assembly of cilia. Dev Cell
2009, 16:867-876.
48. Ikegami K, Heier RL, Taruishi M, Takagi H, Mukai M, Shimma S,
Taira S, Hatanaka K, Morone N, Yao I et al.: Loss of alpha-tubulin
polyglutamylation in ROSA22 mice is associated with
abnormal targeting of KIF1A and modulated synaptic function.
Proc Natl Acad Sci U S A 2007, 104:3213-3218.
49. Lu C, Srayko M, Mains PE: The Caenorhabditis elegans
microtubule-severing complex MEI-1/MEI-2 katanin interacts
differently with two superficially redundant beta-tubulin
isotypes. Mol Biol Cell 2004, 15:142-150.
50. Sharma N, Bryant J, Wloga D, Donaldson R, Davis RC, Jerka-
Dziadosz M, Gaertig J: Katanin regulates dynamics of
microtubules and biogenesis of motile cilia. J Cell Biol 2007,
178:1065-1079.
51. Fukushima N, Furuta D, Hidaka Y, Moriyama R, Tsujiuchi T: Post-
translational modifications of tubulin in the nervous system. J
Neurochem 2009, 109:683-693.
52. Akhmanova A, Steinmetz MO: Tracking the ends: a dynamic
protein network controls the fate of microtubule tips. Nat Rev
Mol Cell Biol 2008, 9:309-322.
www.sciencedirect.com

53. Bieling P, Laan L, Schek H, Munteanu EL, Sandblad L,
Dogterom M, Brunner D, Surrey T: Reconstitution of a
microtubule plus-end tracking system in vitro. Nature 2007,
450:1100-1105.
54. Sandblad L, Busch KE, Tittmann P, Gross H, Brunner D,
Hoenger A: The Schizosaccharomyces pombe EB1 homolog
Mal3p binds and stabilizes the microtubule lattice seam. Cell
2006, 127:1415-1424.
55. Vitre B, Coquelle FM, Heichette C, Garnier C, Chretien D, Arnal I:
 EB1 regulates microtubule dynamics and tubulin sheet
closure in vitro. Nat Cell Biol 2008, 10:415-421.
This paper precisely characterizes the in vitro effects of EB1 at micro-
tubule plus-ends. It shows that EB1 favors lateral association of tubulin to
promote microtubule nucleation and closure. Depending on its concen-
tration, EB1 effect on microtubule structure can increase microtubule
rescue or catastrophe rates.
56. Dixit R, Barnett B, Lazarus JE, Tokito M, Goldman YE,
Holzbaur EL: Microtubule plus-end tracking by CLIP-170
requires EB1. Proc Natl Acad Sci U S A 2009, 106:492-497.
57. Komarova Y, De Groot CO, Grigoriev I, Gouveia SM, Munteanu EL,
 Schober JM, Honnappa S, Buey RM, Hoogenraad CC,
Dogterom M et al.: Mammalian end binding proteins control
persistent microtubule growth. J Cell Biol 2009, 184:691-706.
This in vivo and in vitro study of the role of EB1 in the regulation of
microtubule dynamics shows that EB1 promotes microtubule assembly
and prevents catastrophe in vivo whereas in vitro it can promote cata-
strophe. This report suggests that competition between +TIPs for tubulin
binding at microtubule plus-ends is probably essential for the regulation
of microtubule dynamics.
58. Honnappa S, Gouveia SM, Weisbrich A, Damberger FF,
 Bhavesh NS, Jawhari H, Grigoriev I, van Rijssel FJ, Buey RM,
Lawera A et al.: An EB1-binding motif acts as a microtubule tip
localization signal. Cell 2009, 138:366-376.
This work unravels the existence of a common SxIP EB1-binding motif in
non-CAP-Gly proteins including APC and MCAK. Phosphorylation near
this site may affect EB1 binding and protein recruitment to the plus-ends.
59. Bieling P, Kandels-Lewis S, Telley IA, van Dijk J, Janke C, Surrey T:
 CLIP-170 tracks growing microtubule ends by dynamically
recognizing composite EB1/tubulin-binding sites. J Cell Biol
2008, 183:1223-1233.
This paper describes the mechanism for EB1 autonomous plus-end
tracking activity. EB1 interacts directly with tubulin at the plus-ends of
growing microtubules by recognizing the peculiar structure of the pro-
tofilament.
60. Mishima M, Maesaki R, Kasa M, Watanabe T, Fukata M,
Kaibuchi K, Hakoshima T: Structural basis for tubulin
recognition by cytoplasmic linker protein 170 and its
autoinhibition. Proc Natl Acad Sci U S A 2007, 104:10346-10351.
www.sciencedirect.com
View publication stats
Regulation of microtubule dynamics Etienne-Manneville 111
61. Lee T, Langford KJ, Askham JM, Bruning-Richardson A,
Morrison EE: MCAK associates with EB1. Oncogene 2008,
27:2494-2500.
62. Fong KW, Hau SY, Kho YS, Jia Y, He L, Qi RZ: Interaction of
CDK5RAP2 with EB1 to track growing microtubule tips
and to regulate microtubule dynamics. Mol Biol Cell 2009,
20:3660-3670.
63. Honnappa S, Okhrimenko O, Jaussi R, Jawhari H, Jelesarov I,
Winkler FK, Steinmetz MO: Key interaction modes of dynamic
+TIP networks. Mol Cell 2006, 23:663-671.
64. Kumar P, Lyle KS, Gierke S, Matov A, Danuser G, Wittmann T:
GSK3beta phosphorylation modulates CLASP–microtubule
association and lamella microtubule attachment. J Cell Biol
2009, 184:895-908.
65. Jiang K, Wang J, Liu J, Ward T, Wordeman L, Davidson A, Wang F,
Yao X: TIP150 interacts with and targets MCAK at the
microtubule plus ends. EMBO Rep 2009, 10:857-865.
66. McIntosh JR, Grishchuk EL, Morphew MK, Efremov AK,
Zhudenkov K, Volkov VA, Cheeseman IM, Desai A,
Mastronarde DN, Ataullakhanov FI: Fibrils connect microtubule
tips with kinetochores: a mechanism to couple tubulin
dynamics to chromosome motion. Cell 2008, 135:322-333.
67. Basu R, Chang F: Shaping the actin cytoskeleton using
microtubule tips. Curr Opin Cell Biol 2007, 19:88-94.
68. Broussard JA, Webb DJ, Kaverina I: Asymmetric focal adhesion
disassembly in motile cells. Curr Opin Cell Biol 2008, 20:85-90.
69. Lien WH, Gelfand VI, Vasioukhin V: Alpha-E-catenin binds to
dynamitin and regulates dynactin-mediated intracellular
traffic. J Cell Biol 2008, 183:989-997.
70. Shtutman M, Chausovsky A, Prager-Khoutorsky M,
Schiefermeier N, Boguslavsky S, Kam Z, Fuchs E, Geiger B,
Borisy GG, Bershadsky AD: Signaling function of alpha-catenin
in microtubule regulation. Cell Cycle 2008, 7:2377-2383.
71. Ivanov AI, McCall IC, Babbin B, Samarin SN, Nusrat A, Parkos CA:
Microtubules regulate disassembly of epithelial apical
junctions. BMC Cell Biol 2006, 7:12.
72. Ligon LA, Holzbaur EL: Microtubules tethered at epithelial cell
junctions by dynein facilitate efficient junction assembly.
Traffic 2007, 8:808-819.
73. Shaw RM, Fay AJ, Puthenveedu MA, von Zastrow M, Jan YN,
Jan LY: Microtubule plus-end-tracking proteins target gap
junctions directly from the cell interior to adherens junctions.
Cell 2007, 128:547-560.
Current Opinion in Cell Biology 2010, 22:104–111