Autophagy

ISSN: 1554-8627 (Print) 1554-8635 (Online) Journal homepage: https://www.tandfonline.com/loi/kaup20

DRAM Links Autophagy to p53 and Programmed

Cell Death

Diane Crighton, Simon Wilkinson & Kevin M. Ryan

To cite this article: Diane Crighton, Simon Wilkinson & Kevin M. Ryan (2007) DRAM Links
Autophagy to p53 and Programmed Cell Death, Autophagy, 3:1, 72-74, DOI: 10.4161/auto.3438

To link to this article: https://doi.org/10.4161/auto.3438

Published online: 07 Nov 2006.

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[Autophagy 3:1, 72-74, January/February 2007]; ©2007 Landes Bioscience

DRAM Links Autophagy to p53 and Programmed Cell Death

Addenda

Diane Crighton ABSTRACT

Simon Wilkinson It is clear that changes in autophagy and autophagy regulators occur during tumor

Kevin M. Ryan*
development and that this can have profound effects in certain tumor settings. The fact

.
that p53, a key tumor suppressor mutated in approximately 50% of human cancers, has

E
Tumour Cell Death Laboratory; Beatson Institute for Cancer Research; Cancer now also been shown to induce autophagy, has placed autophagy center stage in the

UT
Research UK Beatson Laboratories; Garscube Estate; Glasgow, UK minds of those interested in the development and treatment of malignant disease. p53 is
*Correspondence to: Kevin M. Ryan; Beatson Institute for Cancer Research;
a transcription factor that responds to cellular stress and prevents the propagation of cells

Garscube Estate; Switchback Road; Glasgow, G61 1BD, UK; Tel.: +441413303655;
Fax: +441419426521; Email: k.ryan@beatson.gla.ac.uk
Received 09/07/06; Accepted 09/28/06
RIB
which may otherwise form a tumor. The recent discovery, therefore, of DRAM (dam-
age-regulated autophagy modulator) as a new p53 target which modulates autophagy is
a major step forward in understanding how p53 controls autophagy and how this relates

IST
to tumor suppression. DRAM is a lysosomal protein that is not only critical for the ability
Previously published online a an Autophagy E-publication: of p53 to induce autophagy, but also for p53’s ability to induce programmed cell death
http://www.landesbioscience.com/journals/autophagy/abstract.php?id=3438

D
- a facet of p53 considered central to its tumor suppressive effects. The fact that DRAM is

KEY WORDS
also inactivated in certain cancers underscores its importance and highlights the possi-

OT
bility that autophagy may have a more profound role in cancer than was first believed.
DRAM, p53, autophagy, apoptosis, cancer

ACKNOWLEDGEMENTS
Work in the Tumor Cell Death Laboratory is
supported by Cancer Research UK. K.R. is a
Cancer Research UK Senior Cancer Research
ON
The number of cellular processes in which autophagy seems to be modulated is ever
increasing.1 The way in which autophagy is controlled in any specific context, however, is
often unclear and requires the identification of signalling pathways that impinge on
.D
autophagy in specific ways. It is now well established that perturbations in autophagy
contribute to tumor development. Core autophagy regulators such as Beclin1 and its
CE

Fellow. binding partner UVRAG are known to be mono-allelically deleted or mutated in certain
tumors and their manipulation in mouse models indicates a causal link between
IEN

Addendum to: autophagy disregulation and cancer.2-8 A number of classical proto-oncoproteins and
tumor suppressors including Akt, PTEN, Ras, p53 and Bcl-2 family members have also
DRAM—A p53-Induced Modulator of Autophagy is Critical
SC

been shown to regulate autophagy.3,9-13 However, for some of these factors, the way in
for Apoptosis which they signal autophagy and the significance in how they control tumor development
BIO

D. Crighton, S. Wilkinson, J. O'Prey, N. Syed, is largely unknown. This is particularly true in the case of p53, but the recent discovery of
P. Smith, P.R. Harrison, M. Gasco, O. Garrone, DRAM (damage-regulated modulator of autophagy) has provided an important link
T. Crook and K.M. Ryan between autophagy and this important tumor suppressor.10
ES

Cell 126:121-34 Human DRAM encodes a 238 amino acid protein that is highly conserved through
evolution with orthologues in insects, fish, worms and amphibian. Interestingly, DRAM
ND

bears no homology to any protein of known function and has no apparent orthologue in
yeast indicating that DRAM may have evolved to control autophagy in a specific context
in higher eukaryotes. DRAM was identified through a screen for genes that are activated
LA

by p53. Moreover, since p53 exerts the majority of effects through transcriptional activa-
tion,14 our studies confirmed that DRAM is a direct target of p53 and that DRAM is
06

induced by chemotherapeutic agents that damage DNA in a p53-dependent manner.

Through a combination of topology predictions, which indicated DRAM has six
20

putative transmembrane regions, and our own immunofluorescence studies, we concluded

that DRAM is localized in the lysosomal membrane. Due to the integral role of lysosomes
©

in the turnover step of autophagy,15 we questioned whether p53 and DRAM regulate
autophagy. Our studies found that p53 and DRAM can induce accumulation of
autophagosomes and that p53 induces successful autophagy (as assessed by studies of
long-lived protein degradation) in a DRAM-dependent manner. At first it seems
counter-intuitive that a protein localizing to lysosomes—which act at a late stage in
autophagy—would be involved in the induction of autophagy. It may be that DRAM
sends a feedback signal to proteins that are involved in the initiation of autophagy—
perhaps by either compromising lysosomal function or other mechanisms. It is also possible
that DRAM works in conjunction with other factor(s) downstream of p53 to bring about

72 Autophagy 2007; Vol. 3 Issue 1

 DRAM Links Autophagy to p53 and Programmed Cell Death
Figure 1. DRAM is required for p53 induced autophagy and programmed
cell death. Cell death via p53, in response to cellular stress, requires
induction of both autophagy via DRAM, and other pro-death signals through
targets such as PUMA, NOXA and BAX to elicit a full cell death response.
Multiple questions remain (dashed grey arrows and text). It is still to be
determined whether other stimuli can induce DRAM e.g., p63/p73 or
pro-autophagic stimuli such as starvation and viral infection. The impact of
mTOR signalling is unclear, and the precise function of DRAM remains
unknown. mTOR, mammalian target of rapamycin; IFN, interferons.
successful autophagy. In this regard, in an independent study,
Feng et al also reported that p53 affects autophagy by modulating
signalling through the mTOR nutrient-sensing kinase which controls
autophagy at an initiation stage.9,16 It is an interesting question,
therefore, whether inhibition of mTOR by p53 is an event upstream
of DRAM function or vice versa or whether the two pathways work
completely independently. Since our studies showed that DRAM
was critical for p53-induced autophagy, this would indicate that the
phosphatidylinositol 3-kinase/mTOR pathway cannot signal
p53-induced autophagy completely independently. However, the
reciprocal experiment to test whether p53’s affects on autophagy are
critically dependent on downregulation of mTOR signalling has not
been addressed and further studies are therefore required.
The ability of p53 to induce programmed cell death has long
been considered critical for its tumor suppressive effects.17 Since
autophagy has been reported to affect cell viability,18 our studies also
examined if DRAM was contributing to cell death regulation.
Although DRAM had no detrimental effects on cell viability when
expressed alone we found that DRAM was highly important for the
ability of p53 to induce programmed cell death. Furthermore, we
found that RNAi knockdown of the ATG5 gene, which encodes a
protein essential for autophagy, also impeded p53-induced cell death
indicating that the effects of DRAM on programmed cell death are
likely through its control of autophagy. Interestingly, we found that
DRAM knockdown could also promote long-term viability following
treatment with an agent that causes p53-dependent DNA damage
responses. This was not the case, however, for knockdown of ATG5.
This therefore hints at an important difference between the point of
action of these two proteins. ATG5 is an ‘effector’ of autophagy
which is required for response to all autophagic stimuli,19 whereas
www.landesbioscience.com Autophagy
DRAM is only a ‘sensor’, signalling autophagy in certain situations,
perhaps to target selective autophagic cargos.20 The further study of
the potential involvement of DRAM function in response to other
stimuli that signal autophagy is required to address this point and is
therefore a worthy avenue of investigation.
Taken together our findings implicate an important role for
DRAM mRNA in tumor suppression. In line with this, we found
DRAM to be downregulated in squamous cancers indicating that there
is a selective pressure to lose DRAM during tumor development. Also,
indicative of an important role of DRAM downstream of p53 we
found this downregulation was often reciprocal to mutation of p53.
This relationship, however, was not completely mutually exclusive
with some tumors exhibiting loss of DRAM expression and mutation
of p53. This indicates that there may be other selective pressures to
lose DRAM due to its tumor suppressive action downstream of
other factors. One obvious possibility is that DRAM is also regulat-
ed by the p53 family members, p63 and p73. p73 in particular has
been implicated in tumor cell death and the response to chemother-
apy21,22 and analysis of whether DRAM plays any role downstream
of p73 is another important question that needs to be addressed. In
summary, therefore, the discovery of DRAM is highly significant in
our quest to understand tumor suppression and tumor cell death.
Clearly, however, in addition to what we have already found, many
important questions still remain (Fig. 1). The further analysis of
DRAM in autophagy and cell death mechanisms as well as for its
potential involvement in the etiology of other tumor types should
prove very rewarding.
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