news and views

Yeast

Human

DNA damage

DNA damage

MEC1

ATM

RAD53

CHK2

DUN1

CRT1

p53

RNR2
RNR3
RNR4

p53R2

Figure 2 Similarities in the DNA damageresponse pathway in yeast and mammals. In

yeast, the protein kinase DUN1 is activated.
Subsequently DUN1 phosphorylates and
inhibits the ability of the CRT1 repressor to
bind to the promoters of several RNR
(ribonucleotide reductase) genes12. (There is also
a weaker CRT1-independent induction pathway,
but it is not as well characterized.) The
activation of DUN1 is controlled by MEC1
(mammalian homologue, ATM/ATR) and
RAD53 (mammalian homologue, CHK2).

DNA after it had been damaged with ultraviolet light or adriamycin. They also showed
that cells in the G1 and G2 phases of the cell
cycle, which do not normally make R2,
induce a DNA-repair capacity in a p53dependent manner, suggesting that p53R2
may be most important for repair in phases
of the cell cycle other than the S phase.
Finally, Tanaka et al. demonstrate that
cells that fail to make p53R2 are more sensitive to killing by DNA-damaging agents.
These overall results could be strengthened
by analysis of cells containing deletions of
p53R2. But even as they stand, the data provide the first direct evidence of a role for p53
in DNA repair and maintenance of genomic
stability.
It has, however, long been appreciated
that dNTPs are involved in DNA repair. Inhibition of ribonucleotide reductase is known
to sensitize cells to DNA damage, and the
transcriptional induction of RNR-encoding
genes occurs in response to DNA damage9,10.
In the yeast Saccharomyces cerevisiae, four
genes encode ribonucleotide reductase and
each is inducible by DNA damage. The DNA
damage-response pathway (also called the
checkpoint pathway) that controls induction of RNR genes is largely conserved with
that in humans which regulates p53. Yeast
have no p53 gene. Instead they have a circuit
in which, in response to DNA damage, the
protein kinase DUN1 is activated. The
details of the pathway are shown in Fig. 2,
along with the mammalian homologues
of the yeast proteins involved. That this
pathway is conserved from yeast to humans
NATURE | VOL 404 | 2 MARCH 2000 | www.nature.com

underscores the importance of dNTP

regulation in cell survival and validates the
relevance of the study of model organisms
to human disease.
These new results1 raise several questions,
the most important of which concerns the
role of p53R2 in tumorigenesis. None of
p53s other transcriptional targets has been
found to be mutated in tumours. Perhaps
this is because p53 activates so many genes
that eliminating just one causes only a slight
increase in tumorigenic potential compared
with eliminating them all at once. But p53R2
may be different. It is directly involved in the
process of repair, and other components of
DNA-repair pathways are known to be
mutated in cancers. For example, mutations
in genes concerned with nucleotide excision
repair cause skin cancer, and mutations in
mismatch repair genes cause colon cancer11.
So a repair gene such as p53R2 may turn out
to be a more important target of p53 for
tumour suppression.
One goal in understanding the role of p53
in tumorigenesis is to uncover a weakness in
p53-mutant tumours that can be exploited
to selectively kill them. Might the lack of
p53R2 induction provide such an opportunity? Because cells lacking p53R2 are sensitive to DNA damage, perhaps what residual
resistance they have is due to dNTPs produced by the cytoplasmic form of ribonucleotide reductase. If one could identify
inhibitors specific for the cytoplasmic form
of the enzyme, these inhibitors might selectively sensitize p53-mutant cells to DNAdamaging chemotherapeutic agents. Specifically inhibiting ribonucleotide reductase

activity in the cytoplasm might allow repair

synthesis to proceed in wild-type cells
through p53R2 expression, while completely
shutting off synthesis in p53-mutant
tumours which would then lack functional
R2 and p53R2 subunits.
To identify such inhibitors it will be necessary to find out which subunits of ribonucleotide reductase combine with p53R2 to
activate dNTP synthesis in the nucleus. Will
it be R1 or an unknown R1-related subunit?
It would be fitting if the most basic of biological processes, dNTP synthesis, proved to be
the Achilles heel of p53-mutant tumours
and most satisfying.

Guillermina Lozano is in the Department of

Molecular Genetics, University of Texas, M. D.
Anderson Cancer Center, Houston, Texas 77030,
USA.
e-mail: gglozano@notes.mdacc.tmc.edu
Stephen J. Elledge is in the Department of
Biochemistry and Molecular Biology, Baylor College
of Medicine, Houston, Texas 77030, USA.
e-mail: selledge@bcm.tmc.edu
1. Tanaka, H. et al. Nature 404, 4249 (2000).
2. Kastan, M. B., Onyekwere, O., Sidransky, D., Vogelstein, B. &
Craig, R. W. Cancer Res. 51, 63046311 (1991).
3. Gottlieb, T. M. & Oren, M. Semin. Cancer Biol. 8, 359368 (1998).
4. Ko, L. J. & Prives, C. Genes Dev. 10, 10541072 (1996).
5. Linke, S. P., Clarkin, K. C. & Wahl, G. M. Cancer Res. 57,
11711179 (1997).
6. Jordan, A. & Reichard, P. Annu. Rev. Biochem. 67, 7198 (1998).
7. Bjorklund, S., Skog, S., Tribukait, B. & Thelander, L.
Biochemistry 29, 54525458 (1990).
8. Engstrom, Y. et al. EMBO J. 3, 863867 (1984).
9. Elledge, S., Zhou, Z. & Allen, J. Trends Biochem. Sci. 17,
119123 (1992).
10. Filatov, D., Bjorklund, S., Johansson, E. & Thelander, L. J. Biol.
Chem. 271, 2369823704 (1996).
11. Lengauer, C., Kinzler, K. W. & Vogelstein, B. Nature 396,
643649 (1998).
12. Huang, M., Zhou, Z. & Elledge, S. J. Cell 94, 595605 (1998).

Circadian clocks

Heartfelt enlightenment
Ueli Schibler
ost animals and plants adapt their
activity to daily cycles of light and
dark. However, their behavioural
and physiological cycles are not merely dictated by changes in light intensity, but are
controlled by an internal timing system
called the circadian clock. As indicated by its
name circa diem means about a day
the circadian clock can measure time only
approximately and must be readjusted every
day by light. So a circadian timing system
needs a light-detector and a way to transfer
the information it detects to the molecular
clockwork1.
How photons entrain biological clocks is
not completely understood, but the paper by
Whitmore et al.2 on page 87 of this issue
describes a startling finding: that circadian
oscillators in zebrafish hearts and kidneys
can be synchronized by light without any

2000 Macmillan Magazines Ltd

input from the eyes. Previously, the same

group showed3 that the expression of Clock,
a master regulatory gene of animal circadian
systems, undergoes daily cycles in many
zebrafish tissues. These include not only
organs known to contain circadian oscillators, such as the pineal gland and the eye, but
also peripheral organs, such as heart, kidney
and spleen. Rhythmic cycles of Clock gene
expression persisted for several days in cultured zebrafish organs, showing that there
are self-sustaining circadian oscillators in
these tissues. Similar observations have also
been made in the fruitfly Drosophila and in
mammals, in which most body cells are
believed to contain circadian clocks4,5.
Whitmore et al.2 noted that the rhythmic
changes in the amounts of Clock messenger
RNA in cultured hearts and kidneys were
greater under a lightdark cycle than in
25

news and views

100 YEARS AGO

A report on the commercial value of the
metric system, with special reference to the
classification of German iron manufactures,
was recently forwarded to the
Wolverhampton Chamber of Commerce by
the British Consul at Amsterdam, and is
referred to in the Board of Trade Journal.
The Consul states that the iron and steel
manufacturers unions of Germany have
adopted a uniform system of dimensions for
articles of universal consumption at home
and abroad. Angle iron of all descriptions,
flanged boiler ends or fronts for Cornish or
Lancashire boilers, the boilers themselves,
and iron and steel tubes and all fittings
connected with them, such as valves, cocks,
T pieces, are made, so far as flange,
diameter, and working lengths are
concerned, in normal standard sizes, in order
that every part of one work may be procured
at once to fit every corresponding part of
another construction At present a
committee of the engineers associations is
occupied in endeavouring to fix a metric
thread for bolts and screws, nuts, bolt-heads,
&c., as the present universal normal standard
(the Whitworth) is so differently constructed
by different works that the parts are not as
interchangeable as should be the case. These
classifications are naturally making more
and more progress in Germany, not in the
iron trade alone, but in other manufactures.
In view of these facts, The Consul points out
that Germany and the Continent generally
will have a constantly increasing advantage
over British manufactures in the future in
foreign countries, unless the metric system
be fully and entirely adopted by Great Britain.
From Nature 1 March 1900.

50 YEARS AGO
Mr. Douglas Kennedy is the director of the
English Folk Dance and Song Society, and as
such knows his subject. He has written a
delightful little book of some 158 pages with
twenty illustrations and two distribution
maps. Dancing throughout the ages has been
for humanity a psychological release and
otherwise. Shortly after the first use of
mustard gas in the First World War, an
American hospital unit took over a complete
British tented hospital in Rouen. The strain of
the personnel was very great; but it was
completely relieved by the introduction of
dances. Dancing was the channel for release
of the overwrought emotion.
From Nature 4 March 1950.
NATURE | VOL 404 | 2 MARCH 2000 | www.nature.com

Drosophila

Zebrafish

Mouse
(and other mammals)

Light

Light

Eye
12
9

Retino-hypothalamic tract

Peripheral clocks

Peripheral clocks
(kidney, heart, etc.)
12

SCN
master clock
Circadian
gene expression

3
6

Chemical signals (?)

12

Peripheral clocks
(kidney, heart, etc.)

3
6

Circadian
gene expression

Figure 1 Synchronization of circadian clocks in cells outside the nervous system. Circadian oscillators
have been found in peripheral cells, such as heart and kidney cells, of insects (Drosophila), fish
(zebrafish) and mammals (rat and mouse). Whitmore et al.2 have now shown that in zebrafish, as in
Drosophila, these oscillators can be synchronized directly by light. In mammals, light entrains master
oscillators in neurons of the suprachiasmatic nucleus (SCN) in the brain, which then synchronize
oscillators in peripheral cells using chemical signals. Like mammals, Drosophila and zebrafish contain
brain structures that are important for circadian behaviour and physiology (lateral pacemaker neurons
in Drosophila, pineal gland in zebrafish). Whether these specialized brain structures contribute to the
entrainment of molecular oscillators in peripheral Drosophila and zebrafish cells is unknown.

constant darkness. They therefore considered that daily light cycles seem to entrain the
oscillations, preventing them from dampening over time. To test this possibility, the
authors monitored Clock gene expression
in heart and kidney organ cultures kept in
reversed lightdark cycles. Two days after
the reversal of the lightdark regimen, the
phase of the Clock messenger RNA cycle had
been offset by 12 hours, showing that heart
and kidney clocks could indeed be entrained
by light.
Even embryonic zebrafish cell lines
can synchronize to lightdark cycles. When
these cells were kept in constant darkness,
2000 Macmillan Magazines Ltd

Clock messenger RNA levels stayed the

same throughout the day. But exposure to
two lightdark cycles caused a significant
daily fluctuation of these transcripts, and
the rhythmic Clock messenger RNA cycle
then persisted for two days under constant
darkness.
Molecular circadian pacemakers consist
of genes specifying proteins that can activate or repress gene transcription, and whose
abundance or activity (or both) can be fine
tuned by other regulatory proteins (such
as protein kinases). The transcriptional activators enhance the expression of the repressors until the latter reach a concentration
27

news and views

sufficient to turn off transcription of their
own genes. The repressor proteins have a
limited lifespan so they eventually decay, and
a new wave of repressor gene transcription
commences. The properties of the system
have evolved to generate oscillations with a
period of about 24 hours6,7. All known components of animal circadian clocks have
homologues in both Drosophila and mouse1.
These include the transcriptional activator
Clock, the putative transcriptional repressors Timeless and Period (three isoforms in
the mouse: mPer1, mPer2 and mPer3), and
blue-light receptors called cryptochromes
(two isoforms in mouse: Cry1 and Cry2)811.
The function of cryptochromes in animal
circadian clocks is enigmatic. In plants and
Drosophila, cryptochromes clearly participate in the light entrainment of circadian
clocks, but in the mouse their function as
light receptors is still controversial8,10,12.
Nonetheless, mouse cryptochromes are
uncontested components of the mammalian
pacemaker, as mutant mice with defects in
both cryptochrome genes, cry1 and cry2, lack
circadian rhythms when kept in constant
darkness and display constitutive expression
of the genes mper1 and mper2 (refs 8,10,13).
The photoreceptors used to entrain peripheral zebrafish clocks are unknown, but Whitmore et al. may be able to use their in vitro
system to test whether cryptochromes are
valid candidates. The recording of action
spectra may be the first step towards this
goal.
In mammals, peripheral clocks are synchronized by different mechanisms from
those in Drosophila and zebrafish (Fig. 1).
Drosophila and zebrafish are both small
and semitransparent, and their peripheral
oscillators can be directly light-entrained by

light receptors2,5,9,11 (such as cryptochromes

in Drosophila). In mammals, which are
opaque, this mechanism is obviously not feasible for setting the time in peripheral oscillators. In these organisms, light adjusts the
circadian pacemaker in part of the brain
called the suprachiasmatic nucleus through
signals from the eyes, and this master pacemaker then synchronizes peripheral clocks
using chemical signals14. Curiously, neither
retinal rods nor cones are required for the
light entrainment of mammalian clocks, so
the light-capturing cells and their photoreceptors remain to be identified15,16.

Ueli Schibler is in the Dpartement de Biologie

Molculaire, Sciences II, Universit de Genve, 30,
Quai Ernest Ansermet, Genve CH-1211,
Switzerland.
e-mail: ueli.schibler@molbio.unige.ch
1. Dunlap, J. C. Cell 96, 271290 (1999).
2. Whitmore, D., Foulkes, N. S. & Sassone-Corsi, P. Nature 404,
8791 (2000).
3. Whitmore, D., Foulkes, N. S., Strahle, U. & Sassone-Corsi, P.
Nature Neurosci. 1, 701707 (1998).
4. Balsalobre, A., Damiola, F. & Schibler, U. Cell 93, 929937
(1998).
5. Plautz, J. D., Kaneko, M., Hall, J. C. & Kay, S. A. Science 278,
16321635 (1997).
6. Rosbash, M. et al. Cold Spring Harb. Symp. Quant. Biol. 61,
265278 (1996).
7. Young, M. W., Wager-Smith, K., Vosshall, L., Saez, L. &
Myers, M. P. Cold Spring Harb. Symp. Quant. Biol. 61,
279284 (1996).
8. Vitaterna, M. H. et al. Proc. Natl Acad. Sci. USA 96,
1211412119 (1999).
9. Stanewsky, R. et al. Cell 95, 681692 (1998).
10. Okamura, H. et al. Science 286, 25312534 (1999).
11. Emery, P., So, W. V., Kaneko, M., Hall, J. C. & Rosbash, M. Cell
95, 669679 (1998).
12. Cashmore, A. R., Jarillo, J. A., Wu, Y. J. & Liu, D. Science 284,
760765 (1999).
13. van der Horst, G.T. et al. Nature 398, 627630 (1999).
14. Brown, S. A. & Schibler, U. Curr. Opin. Genet. Dev. 9, 588594
(1999).
15. Freedman, M. S. et al. Science 284, 502504 (1999).
16. Lucas, R. J. & Foster, R. G. Endocrinology 140, 15201524
(1999).

Physics

Brane new worlds

Jerome Gauntlett
t is common sense that at everyday scales
we live in a world with three large spatial
dimensions. Lisa Randall and Raman Sundrum have recently made the bold suggestion in Physical Review Letters1 that an extra
dimension of infinite extent may supplement the three spatial dimensions we
observe. Not only do they claim that this can
be entirely consistent with current observations, they also argue in a second paper that
closely related scenarios could hold the key
to some fundamental issues in attempts to
unify particle physics with gravity2.
The idea that the four spacetime dimensions that we observe (three space and one
time) could be supplemented by extra
dimensions was first put forward by

28

Theodor Kaluza and subsequently developed by Oskar Klein in the 1920s. The motivation then was to achieve a unification of
Maxwells theory of electromagnetism and
general relativity, which is Einsteins theory
of gravity. They considered an extra fifth
dimension curled up in a small circle that
would be undetected at much longer length
scales. Although Kaluza and Kleins model
turns out to give wrong predictions and so
cannot be a correct description of the real
world, the beautiful idea of extra dimensions
with finite extent (compact dimensions)
has become a central component of string
theory the leading candidate to unify all
of the known forces of nature.
Some of the ingredients in the Randall
2000 Macmillan Magazines Ltd

Gravity

Our world

Hidden world

Figure 1 In brane-world scenarios it is

proposed that our world is a three-brane
propagating within an ambient spacetime of
higher dimension. Particle physics is confined
to the brane and interacts with the extra
dimensions and other hidden three-branes
through gravitational interactions. Randall
and Sundrum1,2 argue that it is possible to
have an extra dimension of infinite extent if
the ambient spacetime is curved in a
particular way.

Sundrum papers were in fact suggested by

developments in string theory. String theory
was originally conceived of as a theory of
oscillating objects with one spatial dimension, or strings. It is now known that in order
for string theory to be mathematically consistent it must also include a rich spectrum
of extended objects of higher dimension:
membranes with two spatial dimensions,
three-branes with three, and so on. Understanding the properties of these branes is
very much an active area of research.
One of the key ideas in the papers by
Randall and Sundrum is that the fourdimensional spacetime we observe at
everyday scales is actually the evolution in
time of a three-brane moving through an
ambient spacetime of higher dimension
(Fig. 1). In such brane-world scenarios,
particle physics is confined to the brane but
the particles can interact with the ambient
spacetime through gravitational interactions. When all the extra spatial dimensions
of the ambient spacetime are compact,
these interactions can be so weak as to have
escaped detection by experiments thus far.
Before the paper by Randall and Sundrum1 it seemed almost obvious that the
ambient spacetime in which the threebrane lives should not be of infinite extent.
In the simplest example with a single, flat,
infinite extra dimension, Newtons law
of gravity would be modified: the force
between two masses on the three-brane
would be inversely proportional to the
cube, not the square, of the distance between
them. The surprising observation made by
Randall and Sundrum is that if the extra
dimension were infinite, and if the embedding spacetime has a very particular curvature, then it is possible for the predicted
NATURE | VOL 404 | 2 MARCH 2000 | www.nature.com