Table Of ContentThe Guanine-Nucleotide
Binding Proteins
Common Structural and
Functional Properties
NATO ASI Series
Advanced Science Institutes Series
A series presenting the results of activities sponsored by the NATO Science Committee,
which aims at the dissemination of advanced scientific and technological knowledge,
with a view to strengthening links between· scientific communities.
The series is published by an international board of publishers in conjunction with the
NATO Scientific Affairs Division
A Life Sciences Plenum Publishing Corporation
B Physics New York and London
C Mathematical Kluwer Academic Publishers
and Physical Sciences Dordrecht, Boston, and London
D Behavioral and Social Sciences
E Applied Sciences
F Computer and Systems Sciences Springer-Verlag
G Ecological Sciences Berlin, Heidelberg, New York, London,
H Cell Biology Paris, and Tokyo
Recent Volumes in this Series
Volume 160-Early Influences Shaping the Individual
edited by Spyros Doxiadis
Volume 161-Research in Congenital HypothyrOidism
edited by F. Delange, D. A. Fisher, and D. Glinoer
Volume 162-Nematode Identification and Expert System Technology
edited by Renaud Fortuner
Volume 163-Leishmaniasis: The Current Status and New Strategies for Control
edited by D. T. Hart
Volume 164-Cochlear Mechanisms: Structure, Function, and Models
edited by J. P. Wilson and D. T. Kemp
Volume 165-The Guanine-Nucleotide Binding Proteins: Common Structural
and Functional Properties
edited by L. Bosch, B. Kraal, and A. Parmeggiani
Volume 166-Vascular Dynamics: Physiological Perspectives
edited by N. Westerhof and D. R. Gross
Series A: Life Sciences
The Guanine-Nucleotide
Binding Proteins
Common Structural and
Functional Properties
Edited by
L. Bosch and B. Kraal
Leiden University
Leiden, The Netherlands
and
A. Parmeggiani
Ecole Poly technique
Palaiseau, France
Springer Science+Business Media, LLC
Proceedings of an EMBO-NATO-CEC Advanced Research Workshop
on the Guanine-Nucleotide Binding Proteins:
Common Structural and Functional Properties,
held August 6-11, 1988,
in Renesse, The Netherlands
Library of Congress Cataloging in Publication Data
EMBO-NATO-CEC Advanced Research Workshop on the Guanine-Nucleotide
Binding Proteins: Common Structural and Functional Properties (1988: Renesse,
Netherlands)
The guanine-nucleotide binding proteins: common structural and functional
properties I edited by L. Bosch and B. Kraal and A. Parmeggiani.
p. cm.-(NATO ASI series. Series A, Life sciences; v. 165)
"Proceedings of an EMBO-NATO-CEC Advanced Research Workshop on the
Guanine-Nucleotide Binding Proteins: Common Structural and Functional Pro
perties, held August 6-11,1988 in Renesse, The Netherlands"-T.p. verso.
"Published in cooperation with NATO Scientific Affairs Division."
Includes bibliographical references and indexes.
ISBN 978-1-4757-2039-6 ISBN 978-1-4757-2037-2 (eBook)
DOI 10.1007/978-1-4757-2037-2
1. G proteins-Structure-activity relationships-Congresses. I. Bosch, L. II.
Kraal, B. III. Parmeggiani, A. IV. North Atlantic Treaty Organization. Scientific Af
fairs Division. V. Title. VI. Series.
QP552.G16E52 1988 89-4038
574.19'245-dc20 CIP
© 1989 Springer Science+Business Media New York
Originally published by Plenum Press, New York in 1989.
Softcover reprint of the hardcover 1st edition 1989
All rights reserved
No part of this book may be reproduced, stored in a retrieval system, or transmitted
in any form or by any means, electronic, mechanical, photocopying, microfilming,
recording, or otherwise, without written permission from the Publisher
PREFACE
This volume contains the proceedings of the EMBO-NATO-CEC Advanced
Research Workshop on "Guanine-nucleotide binding proteins. Common
structural and functional properties", which was held in Renesse, The
Netherlands, August 6-11, 1988.
The transmission of information is one of the most important
processes in cellular life and involves the most diverse physiological
functions. The cellular membrane, as the obligatory target for external
signals, harbours complex pathways transducing the signals from the
receptors of the external stimuli to the cytoplasmic effector.
Heterotrimeric proteins are fundamental components of these pathways.
Other proteins that are monomeric may be found associated with the
membrane or in soluble form in the cytoplasm, and' can also function in
signal transduction. Intracellular transmission of signals may proceed in
an analogous fashion, protein synthesis being a well-known example.
It is one of the most remarkable and puzzling observations of recent years
that all of these proteins share common properties, both functionally and
structurally. The ir primary structures show pronounced s imilari ties, in
most cases concentrated in the NH2-terminal portion of the molecule. They
all bind guanine nucleotides (hence the general name of G-proteins) and
are GTPases, a crucial enzymatic activity, since it converts the active
complex induced by GTP into the inactive one induced by GDP. Consensus
sequences have been ident if ied as responsable for interact ing with the
different parts of the guanine nucleotide.
To emphasize the importance of this class of proteins in the pathology of
mammals, it is sufficient to mention that C\lteration of cell
prol iferat ion, such as oncogenic transformat ion, is assoc iated with the
mutation of the ras product p21, a guanine-nucleotide binding protein.
For many years there has been little, if any, interaction between the
groups of scientists involved in the study of these various protein
famil ies. The exponent ial increase of knowledge of the ir funct ioning and
the growing not ion of mechanist ic similar i ties have led to the
organization of the above-mentioned workshop. Its major goal was to bring
together a number of specialists, actively engaged in the study of one or
more of the major families of these proteins, in order to stimulate the
exchange of information and to coordinate the efforts.
For four and a half day, similarities and diversities of these proteins
were discussed, taking advantage of the detailed analysis of two model
proteins: the prokaryotic elongation factor Tu and the ras protein p21.
They are the only members of these families that have been crystallized,
which has led to the elucidation of their three-dimensional structures.
The workshop was highly favoured by the relaxing atmosphere in a
comfortable environment far away from the tumult of daily life, and the
salted sea breeze of Zeeland. It could not have been organized without the
v
generous support from the European Molecular Biology Organization, from
the NATO Scientific Affairs Division, and from the Commission of the
European Communities. Financial contributions from the Royal Netherlands
Academy of Arts and Sciences, Gist Brocades NV, Rhone-Poulenc Sante, and
Du Pont de Nemours (Nederland) BV are also gratefully acknowledged.
We have the feeling that everyone participated intensively and that
the exchange of information was useful and exciting. We hope that any of
the stimulating propositions will lead to further investigations and the
future development of the field. If so, this will be the most appreciated
reward for the effort of the organizers.
L. Bosch,
B. Kraal,
A. Parmeggiani
vi
CONTENTS
THREE-DIMENSIONAL STRUCTURES AND CONSENSUS SEQUENCE ELEMENTS
New Structural Data on Elongation Factor-Tu:GDP Based on
X-ray Crystallography . . . . . . . 3
J. Nyborg and T. la Cour
Progress on the Three-Dimensional Structural Determination of
Trypsin-Modified EF-Tu-GDP . . . . . . . . 15
F. Jurnak, M. Nelson, M. Yoder, S. Heffron, and S. Miu
Three-Dimensional Structure of ras p21 Proteins . . 27
A.M. de Vos, L. Tong, M.V. Milburn, P.M. Matias, and S.-H.
Kim
The GTP-Binding Domain Revisited 35
Th.E. Dever and W.C. Merrick
STRUCTURE, FUNCTION AND GENETICS OF TRANSLATIONAL FACTORS
Novel Mutants of EF-Tu 51
D. Hughes and C.G. Kurland
The Elongation Factor EF-Tu from E. coli Activates the tRNA
tufB Operon in Trans by Binding to a Cis-Acting Region
Upstream of the Promoter . . . . . . . . 57
E. Vijgenboom, L. Nilsson, A. Talens, and L. Bosch
Structure-Function Relationships of the GTP-Binding Domain of
Elongation Factor Tu . . . . . .. ..... . 67
P.H. Anborgh, R.H. Cool, E. Jacquet, M. Jensen, G.
Parlato, and A. Parmeggiani
A Mutation that Hinders the GTP Induced Aminoacyl-tRNA
Binding of Elongation Factor Tu 77
Y.-W. Hwang, F. Jurnak and D.L. Miller
The Application of Fluorescent and Photosensitive Analogues
of Guanine Nucleotides to the Function and Structure of
G-Binding Proteins . . . . . . . . . . . . . . . . . . 87
J.F. Eccleston, T.F. Kanagasabai, D.P. Molloy, S.E. Neal,
and M.R. Webb
Affinity Labeling of the GDP/GTP Binding Site in Thermus
thermophilus Elongation Factor Tu . . . . . . . . . 99
M.E. Peter and M. Sprinzl
Characterization of Elongation Factor Tu from Bacillus
subtilis Modified by Affinity Labeling ..... III
J. Jonak, K. Karas, and!. Rychlik
Effects of Kirromycin on the Elongation Factor EF-Tu and its
Interactions with GDP or GTP and tRNA. The Application
of Zone-Interference Gel Electrophoresis, a New Method
for the Analysis of Weak Complexes . . . . . . . . . . 121
B. Kraal, J.P. Abrahams, and L. Bosch
Factors and Ribosomes: Their coupling and Mode of Signal
Processing . . . . .. . ...... . 131
W. Moller and R. Amons
The Structure and Regulation of Mammalian Initiation
Factor eIF2 . . . . . . . . . . . . . . . . . . 143
J.W.B. Hershey, V.K. Pathak, H. Ernst, M. Humbelin, and
R.J. Kaufman
STRUCTURE, FUNCTION AND GENETICS OF ras PROTEINS
Structure of the Human ras Gene Family . . . . . . . . . . . 153
P. Chardin, N. Touchot, A. Zahraoui, V. Pizon, I. Lerosey,
B. Olofsson, and A. Tavitian
A Mutational Analysis of ras Function . . . . . . . . . 165
B.M. Willumsen, H. Adari, K. Zhang, A.G. Papageorge,
J.C. Stone, F. McCormick, and D.R. Lowy
Analysis of the Biochemical and Biological Activities of
Deletion Mutants of the H-ras p21 Protein Suggest that
GAP is an Essential Component of its Effector Function 179
A. Di Donato, S.K. Srivastava, and J.C. Lacal
Purification and Molecular Cloning of Bovine GAP ..... . 191
J.B. Gibbs, U.S. Vogel, M.D. Schaber, M.S. Marshall, R.E.
Diehl, E.M. Scolnick, R.A.F. Dixon, and I.S. Sigal
The Function of the Mammalian ras Proteins ..... 201
A. Hall, J.D.H. Morris, B. Price, J.F. Hancock, S.
Gardener, M.D. Houslay, M.J.O. Wakelam, and C.J.
Marshall
Biochemical Properties of Ha-ras Encoded p21 Mutants 209
J. John, M. Frech, J. Feuerstein, R.S. Goody, and F.
Wittinghofer
Structural and Functional Studies on c-p21, v-p21 and the and
Genetically Engineered Guanine Nucleotide Binding
Domain of EF-Tu . . . . . . . . . . . . . . . . . . 215
A. Pingoud, U. Pieper, R. Busche, H.-J. Ehbrecht, M.
Wehrmann, F.-U. Gast, J. Feuerstein, A. Wittinghofer,
Th. Jarchau, G.-W. Kohring, and F. Mayer
Structure and Function of ras p21: Studies by Site-Directed
Mutagenesis . . . . . . . . . . . . . . . . . . . . . 229
Th.Y. Shih, D.J. Clanton, P. Saikumar, L.S. Ulsh, and S.
Hattori
viii
The Cyclic AMP Producing Pathway in Saccharomyces cerevisiae
Involves CDC25 and RAS Genes Products 241
M. Jacquet, J. Camonis, E. Boy-Marcotte, F. Damak, and H.
Garreau
Mutations at the RAS2 Locus that, in a rasl- Background,
Impair the Growth of Yeast on Nonfermentable Carbon
Sources . . . . . . . . . . . . . .... 251
O. Fasano, J.B. Crechet, E. De Vendittis, R. Zahn,
G. Feger, A. Vitelli, and A. Parmeggiani
Structural and Functional Analysis of ypt Proteins, a Family
of ras-Related Nucleotide-Binding Proteins in
Eukaryotic Cells . . . . . . . . . . . . . . 257
D. Gallwitz, H. Haubruck, C. Molenaar, R. Prange, M.
Puzicha, H.D. Schmitt, C. Vorgias, and P. Wagner
Analysis of the ras Gene Function in Dictyostelium
discoideum .......... ..... . 265
C.D. Reymond, M.E.E. Luderus, G.N. Europe-Finner, N.A.
Thompson, E. Burki, R. Van Driel, and P.C. Newell
Identification of Guanine-Nucleotide Binding Proteins in
Plants: Structural Analysis and Evolutionary Comparison
of the ras-Related ypt-Gene Family from Zea mays 273
K. Palme, Th. Diefenthal, C. Sander, M. Vingron, and J.
Schell
Does HIV neE Protein Belong to the G-Protein Family? . . . . 285
B. Guy, M.P. Kieny, Y. Riviere, M. Girard, L. Montagnier,
and J.P. Lecocq
STRUCTURE, FUNCTION AND GENETICS OF SIGNAL TRANSDUCING PROTEINS
Structure and Function of G Proteins from Mammalian and Yeast
Cells . . . . . .. ........... .
291
Y. Kaziro
Three Forms of Gi Discriminated by Synthetic Peptide Antisera
S.M. Mumby. . . . . . . . . . . .. .... . ...
305
Identification of Functional Domains in G Protein a Chains 313
S.B. Masters, R.T. Miller, K.A. Sullivan, and H.R. Bourne
Behavior of Membrane Guanine-Nucleotide Binding Proteins as
Signal Transducers . . . . . . . . . . . . . . . . . 325
M. Ui and T. Katada
Transducin: the Molecular Switch in Visual Excitation and a
Model for Biological Coupling Enzymes . . . . . 337
Y.-K. Ho and V.N. Hingorani
A Novel Cytosolic GTP-Binding Protein with Phospholipid
Stimulated GTP-Binding and GTPase Activity 349
R. Sagi-Eisenberg, L.M. Traub, G. Gat-Yablonski, and M.
Aridor
Role of a G Protein Homolog in Yeast Pheromone Response . . . . .. 359
J. Kurjan
Signal-Transducing G-Proteins in Dictyostelium discoideum . . 369
A.A. Bominaar, B.E. Snaar-Jagalska, F. Kesbeke, and P.J.M.
Van Haastert
STRUCTURE AND FUNCTION OF TUBULINS
Tubulin as a G-Protein: Regulation of Tubulin-Tubulin
Interactions by GTP Hydrolysis . . . . . . . .
379
M.-F. Carlier and D. Pantaloni
Tubulin Structure and Nucleotide Binding 385
E.-M. Mandelkow, K. Linse, and E. Mandelkow
Tubulin as a G Protein? . 391
M.M. Rasenick, Y. Kun, and N. Wang
Author Index 403
Subject Index 405
x
