Regulation of the RAS Signaling Network

209 Pages·1996·4.76 MB·English

by  Hiroshi Maruta, Antony W. Burgess (auth.)

MOLECULAR
BIOLOGY
INTELLIGENCE
UNIT
REGULATION OF THE
RAS SIGNALING
NETWORK
Hiroshi Maruta
Antony W. Burgess
Ludwig Institute for Cancer Research
Royal Melbourne Hospital
Victoria, Australia
CHAPMAN & HALL
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MOLECULAR BIOLOGY INTELLIGENCE UNIT
REGULATION OF THE RAS SIGNALING NETWORK
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ISBN-13: 978-1-4612-8502-1 e-ISBN-13: 978-1-4613-1183-6
DOl: 10.1007/978-1-4613-1183-6
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Library of Congress Cataloging-in-Publication Data
Regulation of the RAS signaling network / [edited by} Hiroshi Maruta,
A.W. Burgess
p. cm. - (Molecular biology intelligence unit)
Includes bibliographical references and index.
1. Ras oncogenes. 2. Cellular signal transduction I. Maruta, Hiroshi. II. Burgess,
Antony, 1946- . III. Series.
RC268.44.R37R44 1996
616.99'4071-dc20
96-35747
CIP
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IN MEMORY OF IRVING SIGAL
M
uch of the study of the molecular
biology ofRAS-in particular, the
structure-function work-was initiated
by the late Irving Sigal (1953-1988),
when he joined Ed Scolnick's group at
Merck, Sharp & Dohme Research
Laboratories in 1983. Irving was a
chemist by training. Using site-directed
mutagenesis to create a series of RAS
Irving Sigal (7953-1988)
mutants, he and his colleagues identi-
fied the unique domain of RAS which is required for its interaction
with downstream molecules (for further details, see chapter 5). This
section of RAS is now called the "effector" or "switch I" domain. The
"effector" domain mutants of RAS, many of which are no longer
oncogenic, have served as useful tools for distinguishing between the
mode of interaction between RAS and many RAS-binding proteins.
Irving was also actively involved in both the purification and cloning
of the first RAS effector protein, called GAP 1, a RAS GAP of 120
kDa (see chapter 5). If Irving were still alive, it is most likely that he
would have joined us in editing this first book on RAS.
Unfortunately, soon after the cloning of GAP-1 was completed
PanAm flight 103, carrying 270 people including Irving, was downed
over Lockerbie in England by an act of terrorism on December 21,
1988. There were, sadly, no survivors of this tragic air crash. It was a
great loss not only for his own family, in particular his wife Cathy
Sigal, but also for us and the whole RAS research community. Thus,
we would like to dedicate our first RAS book to the late Irving Sigal.
Irving was a great seeder of the RAS field, and, as will be seen in this
book, the world is now harvesting fruit from the rich plantation that
he left us. We hope that this book will also stimulate others to creative
efforts in the field of RAS research, bringing new fruits to both this
and coming generations ...
CO NTE NTS
r;::::================== ================~
1. Genetics of RAS Signaling in Drosophila .................................... 1
David D. L. Bowtell
I. Introduction ................................................................................... 1
II. Background to the Experimental Systems ....................................... 2
III. Protein Tyrosine Kinase Receptors ................................................. 6
IV. Linking RAS with Tyrosine Kinase Receptors .............................. 14
V. A Kinase Cascade Downstream of RAS ........................................ 20
VI. Nuclear Events ............................................................................. 25
VII.Concluding Remarks .................................................................... 34
2. RAS-Mediated Signal Transduction in C. elegans ..................... 47
Min Han and Meera Sundaram
I. Introduction ................................................................................. 47
II. Genetic Approaches Used to Study Vulval Signal Transduction ... 50
III. Genetic and Molecular Analysis of the let-GO RAS Gene ............... 57
IV. Genes Acting Upstream ofRAS .................................................... 59
V. Genes Acting Downstream of RAS ............................................... 65
VI. Multiple Functions of the let-GO RAS-mediated Signal
Transduction Pathway During C. elegans Development ................ 68
3. Mammals I: Regulation of RAS Activation ............................... 75
Antony W Burgess
I. Introduction ................................................................................. 75
II. Growth FactoriCytokine Signaling ............................................... 77
III. Growth Factor Cascades: Autocrine Stimulation .......................... 85
4. Prenylation ofRAS and Inhibitors ofPrenyltransferases .......... 95
Isabel Sattler and Fuyuhiko Tamanoi
I. Introduction ................................................................................. 95
II. C-Terminal Modification of RAS Proteins ................................... 96
III. Farnesyltransferase and Geranylgeranyltransferases ....................... 98
IV. Mutational Analyses ofPrenyltransferases ................................... 103
V. Inhibitors of Prenylation ............................................................ 105
VI. Biological Effects ofPrenyltransferase Inhibitors ......................... 120
5. Mammals II: Downstream ofRAS and Actin-Cytoskeleton .... 139
Hiroshi Maruta
I. RAS GAPs .................................................................................. 139
II. Oncogenic Mutations ofRAS ..................................................... 143
III. Effectors ofRAS ......................................................................... 145
IV. Actin-Cytoskeleton ..................................................................... 159
V. G Proteins in the Rho Family (Rho, Rac and CDC42) .............. 162
VI. RAS-Activated or -Repressed Genes ............................................ 164
6. From RAS to MAPK: Cell-Free Assay System for
RAS- and Rapl-Dependent B-Raf Activation ......................... 181
Kazuya Shimizu, Toshihisa Ohtsuka and Yoshimi Takai
I. Introduction ............................................................................... 181
II. Cell-free Assay System for the RAS-dependent Activation
of the MAP Kinase Cascade in Xenopus Oocyte Cytosol ............ 182
III. REKS ......................................................................................... 184
IV. Raf Activation ............................................................................ 188
V. Conclusion ................................................................................. 193
Index .............................................................................................. 201
r.======================E DI T O RS
=======================::;-]
Hiroshi Maruta
Ludwig Institute for Cancer Research
Royal Melbourne Hospital
Victoria, Australia
Chapter 5
Antony W. Burgess
Ludwig Institute for Cancer Research
Royal Melbourne Hospital
Melbourne, Australia
Chapter 3
t==================== CO NT RI BU TO RS=========I
David D.L. Bowtell Kazuya Shimizu
Peter MacCallum Cancer Institute Department of Molecular Biology
Melbourne, Australia and Biochemistry
Chapter 1 Osaka University Medical School
Suita, Japan
Min Han Chapter 6
Department of Molecular, Cellular
and Developmental Biology Meera Sundaram
University of Colorado Department of Molecular, Cellular
Boulder, Colorado, U.S.A. and Developmental Biology
Chapter 2 University of Colorado
Boulder, Colorado, U.S.A.
Toshihisa Ohtsuka Chapter 2
Department of Molecular Biology
and Biochemistry y oshimi T akai
Osaka University Medical School Department of Molecular Biology
Suita, Japan and Biochemistry
Chapter 6 Osaka University Medical School
Suita, Japan
Isabel Sattler Chapter 6
Department of Microbiology
and Molecular Genetics Fuyuhiko Tamanoi
University of California Department of Microbiology
Los Angeles, California, U.S.A. and Molecular Genetics
Chapter 4 University of California
Los Angeles, California, U.S.A.
Chapter 4
===================== PREFACE =====================
C
ancer research has revolutionized our understanding of mammalian cell
biology, and in many respects the discoveries associated with the RAS
protein have been pivotal to our spectacular progress. A little over thirty years
ago, Jennifer Harvey discovered that a new sarcoma virus had been produced by
JJ,
passaging the murine Moloney Leukemia virus in rats (Harvey, Nature
204: 11 04, 1964). The new virus (Ha-MSV) rapidly induced sarcomas near the
site of injection of the virus. Eventually, Edward Scolnick and his team at the
National Cancer Institute in Bethesda determined that Ha-MSV contained a
large insert of rat genetic material between the Moloney helper virus sequences
(Scolnick EM and Parks WP, J Viro113: 1211, 1974). Clearly, the rat sequences
were critical for the acute-transforming properties of the new virus (Shih TY,
Weeks MO, Young HA and Scolnick EM, J ViroI31:546, 1979). Soon after the
RAS studies were published, Michael Bishop, Harold Varmus and Dominique
Stehelin determined that another viral oncogene, Src, had also been captured
from its host cells (Stehelin E et aI, Nature 260:170, 1976).
Although the link between human cancers and the oncogenic animal viruses
had been difficult to detect, Robert Weinberg and his colleagues believed that
the genetic information associated with human cancers would also be capable of
transforming normal cells. They adapted a DNA transfection technique to
detect the cancerous gene(s) associated with human tumors by focus formation
in a mouse fibroblast cell line (Shih C, Shilo BZ, Goldfarb MP, Dannenberg A
and Weinberg RA. Proc Natl Acad Sci 76:5714, 1979). Almost simultaneously
three independent groups determined that the human oncogene identified by
Weinberg's team corresponded to the rat gene (RAS) responsible for the trans
forming properties of the Harvey and Kirsten sarcoma viruses (Shih et al, Nature
290:261, 1981; Krontiris TG and Cooper GM Proc Natl Acad Sci 78:1181C,
1981; Perucho M et al Cell 27:467, 1981). Clearly, some of the mutations
responsible for human cancer operated by mechanisms similar to the oncogenes
carried by cancer-causing viruses of animals.
As well as identifying a myriad of possible mutations which can contribute to
the cancerous state, work over the last fifteen years has also confirmed the
commonality of many oncogenic events (e.g., p53 and RAS). Despite the
enormous diversity of this research, the centrality of the RAS oncogene and the
c-RAS protein in cellular proliferative and differentiative processes has remained.
The initial observations indicated that the oncogenic RAS and normal
human RAS proteins differed by a single amino acid. Scolnick's group had
observed that the viral RAS (v-RAS) protein was phosphorylated and was
capable of binding GTP (Shih TY et al, J. Virol42:253, 1982). Although normal
forms of human RAS bind GTP with equal affinity, normal RAS hydrolyzes the
GTP to GDP at a much higher rate than the oncogenic RAS (McGrath et aI,
Nature 310:644, 1984). More importantly, the intrinsic GTPase activity of only
normal RAS, and not oncogenic RAS, is highly stimulated by GTPase activating
proteins (GAPs) (Trahey M and McCormick F, Science 238:542, 1987). By
analogy with other G-protein signaling systems, it was suggested that only the
GTP-bound form of RAS acted to signal mitogenic, differentiative or motility
processes. It is now clear that many cellular regulatory systems act by controlling
the concentration of membrane-associated GTP-RAS.
The importance of RAS continues to grow as the biochemistry of more and
more cytokine/growth factorltransforming systems are unraveled. We now have
exquisite details of the three-dimensional structure of RAS (Franken SM et aI,
Biochemistry 32:8411, 1993; Milburn MV et aI, Science 297:939, 1990), and
there is even a high resolution crystal structure of the RAS/Raf interface (Nassar N
et aI, Nature 375:1554, 1995). It is clear that activated RAS controls several
critical processes, but whilst the roles of these processes in normal cell division or
the maintenance of the oncogenic state remain to be determined, oncogenic RAS
is clearly an important target for the development of more specific and potent
cancer therapies Qames GL et aI, Science 260:1937,1993;), (Sepp-Lorenzino L
et al, Cancer Res 55:5302, 1995).
Tony Burgess
Hiroshi Maruta
May, 1996
=================== CH APTE R1 = =================
GENETICS OF RAS SIGNALING
IN DROSOPHILA
David D.L. Bowtell
I. INTRODUCTION
A
s in other multicellular organisms, RAS proteins playa key role in
the signaling pathways that regulate cell growth and differentiation
in Drosophila. Although these proteins were first identified in Droso
phila using cross-species hybridization, it has been the use of classical
genetics which has overwhelmingly driven investigations of RAS func
tion in Drosophila. This focus has been successful in placing RAS in
complex signaling pathways for several reasons: genetic screens gener
ally make no assumptions about the biochemical activity of the com
ponents of the signaling pathway, thereby facilitating the isolation of
proteins with novel activities; a functional requirement for the protein
in the pathway is established at the outset, which is often lacking when
proteins are isolated by other means; and tests of cell autonomy and
epistasis frequently position the requirement for the protein in the path
way, even before its biochemical activity is known.
Although RAS proteins are involved in many facets of Drosophila
development, this review will focus mainly on their role in the signal
ing pathways downstream of three tyrosine kinase receptors; sevenless,
the Drosophila EGF receptor homologue (D-EGFR) and torso. Sevenless
has a very restricted role in development, being specifically required
for differentiation of the R7 photoreceptor cell in the developing eye.
Torso is required in several sites but its role in the development of
the terminal structures of the embryo is best characterized. The 0-EGFR
is required in many tissues, including the development of the embry
onic ventro-Iateral midline cells, the adult cuticle, wings and photore
ceptor cells.
Regulation of the RAS Signaling Network, edited by Hiroshi Maruta
and Antony W. Burgess. © 1996 R.G. Landes Company.