Table Of Content286
Current Topics
in Microbiology
and Immunology
Editors
R.W. Compans, Atlanta/Georgia
M.D. Cooper, Birmingham/Alabama
T. Honjo, Kyoto· H. Koprowski, Philadelphia/Pennsylvania
F. Melchers, Basel· M.B.A. Oldstone, La Jolla/California
S. Olsnes, Oslo· M. Potter, Bethesda/Maryland
P.K. Vogt, La Jolla/California· H. Wagner, Munich
Springer-Verlag Berlin Heidelberg GmbH
I.H. Madshus (Ed.)
Signalling from
Internalized
Growth Factor
Receptors
With 19 Figures
Springer
Professor Inger Helene Madshus, M.D., Ph.D.
Institute of Pathology
Rikshospitalet University Hospital
0027 Oslo
Norway
e-mail: [email protected]
Cover illustration by LH. Madshus
The receptor tyrosine kinase ErbB2 (green) is over-expressed in the breast carcinoma
cell line SKBr3. ErbB2 is endocytosis deficient and excluded from early endosome
antigen positive endosomes (blue).
ISSN 0070-217X
ISBN 978-3-642-05912-4 ISBN 978-3-540-69494-6 (eBook)
DOI 10.1007/978-3-540-69494-6
Library of Congress Catalog Card Number 72-152360
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© Springer-Verlag Berlin Heidelberg 2004
Originally published by Springer-Verlag Berlin Heidelberg New York in 2004
Softcover reprint of the hardcover I st edition 2004
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Preface
Mammalian cells are to a large extent controlled by the environment. Dif
fusible factors (growth factors, cytokines, and hormones) released by oth
er cells in the body bind to and activate receptors localized at the cell sur
face. In the case of the fibroblast growth factor receptor, there seems to be
receptors both at the plasma membrane and in the nucleus. Cellular recep
tors control growth, apoptosis, immune function, differentiation, develop
ment, and upon dysregulation, cancer progression and metastasis. Upon li
gand binding, most receptors are internalized. However, the mechanisms
of endocytosis are diverse, and receptors are taken into cells from different
membrane microdomains. Activation of receptors results in two important
interconnected processes, namely, signal transduction and endocytosis. In
terestingly, signal transduction controls endocytosis and endocytosis con
trols signalling. In both processes sequential formation of transient protein
machineries is crucial. Currently, characterization of such complex ma
chineries is advancing rapidly. It has recently become appreciated that sev
eral post-translational modifications directly control the affinity of pro
tein-protein interactions. This volume of Current Topics in Microbiology
and Immunology focuses on the recent understanding of signalling from in
ternalized activated growth factor receptors. This includes information on
pathways by which the rate and specificity of endocytosis and intracellular
sorting are controlled. It further includes information on how specialized
signalling and trafficking platforms are formed at the plasma membrane
and on intracellular vesicles. Recent advances in cell biology and bioinfor
matics have revealed the existence of several conserved protein modules,
such as UIM, UEV, UBA, and CUE domains, that endow proteins with the
ability to bind the small, conserved peptide ubiquitin. Ubiquitination
thereby turns out to be a key process in controlling affinity and specificity
of protein interactions and therefore in controlling both signal transduc
tion and intracellular transport. As discussed in the chapter by Sigismund
et aI., evidence is accumulating that monoubiquitination, in addition to
controlling membrane trafficking of receptors, controls histone function,
transcription regulation, DNA repair, and DNA replication. This opens up
a new and exciting avenue in science that will eventually shed light on
some of the fundamental and complex processes of cell biology.
Inger Helene Madshus
List of Contents
Receptor Tyrosine Kinase Signaling and Trafficking
Paradigms Revisited
M.A. Barbieri, T.P. Ramkumar, S. Fernadez-Pol, P.I. Chen,
and. P.D. Stahl. ............................................ .
Met Receptor Dynamics and Signalling
D.E. Hammond, S. Carter, and M.J. Clague 21
Signaling, Internalization, and Intracellular Activity
of Fibroblast Growth Factor
A. Wifdlocha and V. S0rensen. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45
Ubiquitin System-Dependent Regulation
of Growth Hormone Receptor Signal Transduction
G.J. Strous, C. Alves dos Santos, J. Gent, R. Govers, M. Sachse,
J. Schant!, and P. van Kerkhof . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 81
Clathrin-Independent Endocytosis and Signalling
of Interleukin 2 Receptors
IL-2R Endocytosis and Signalling
F. Gesbert, N. Sauvonnet, and A. Dautry-Varsat. . . . . . . . . . . . . . . . . .. 119
Signaling Through Monoubiquitination
S. Sigismund, S. Polo, and P.P. Di Fiore ......................... 149
Subject Index ............................................. 187
List of Contributors
(Their addresses can be found at the beginning of their respective chapters.)
Alves dos Santos, C. 81 Kerkhof, P. van 81
Barbieri, M.A. Polo, S. 149
Carter, S. 21 Ramkumar, T.P. 1
Chen, P.I. 1 Sachse, M. 81
Clague, M.J. 21 Sauvonnet, N. 119
Dautry-Varsat, A. 119 Schantl, J. 81
Di Fiore, P.P. 149 Sigismund, S. 149
Fernadez-Pol, S. S0rensen, V. 45
Gent, J. 81 Stahl, P.D.
Gesbert, F. 119 Strous, G.J. 81
Govers, R. 81 Wicrdlocha, A. 45
Hammond, D.E. 21
CTMI (2004) 286:1-19
© Springer-Verlag 2004
Receptor Tyrosine Kinase Signaling
and Trafficking-Paradigms Revisited
M. A. Barbieri· T. P. Ramkumar· S. Fernadez-Pol· P. 1. Chen·
P. D. Stahl (~)
Department of Cell Biology and Physiology, School of Medicine,
Washington University, 660 S. Euclid Avenue, Campus Box 8228, St. Louis,
MO 63110, USA
pstahl@cellbiology. wustl.edu
Introduction . . . . . . . . . . . . .
2 Endocytosis of Signaling Receptor . 2
3 Receptor Trafficking: Endocytic Regulation of Signaling Pathways . 4
4 Signaling in Endosomes . 7
5 Sorting in Endosomes . . 11
6 A Perspective on Signaling Endosomes 13
References. . . . . . . . . . . . . . . . . . . . . 14
Abstract The recognition of growth factors and other cell signaling agents by their
cognate cell surface receptors triggers a cascade of signal transducing events. Ligand
binding and subsequent activation of many signal transducing receptors increases
their rate of internalization. Endocytosis of the receptor has always been viewed as
primarily a mechanism for signal attenuation and receptor degradation, but recent
evidence suggests that internalization may result in the formation of specialized sig
naling platforms on intracellular vesicles. Thus, understanding how interactions be
tween receptors and intracellular signaling molecules, such as adaptors, GTPases,
and kinases, are regulated will undoubtedly provide insight into the ways that cells
sense and adapt to the extracellular milieu.
1
Introduction
Signaling from the extracellular environment is achieved through un
ique, dynamic, and efficient signal transduction systems. The first cellu
lar components that come in contact with external signals are cell sur
face receptors. Receptor tyrosine kinases (RTKs) constitute a large
2 M.A. Barbieri et al.
group of receptors that respond to growth factors and have intrin
sic tyrosine kinase activity. On ligand binding, RTKs dimerize and,
through a conformation change, intrinsic cytoplasmic kinase activity is
switched on, which in turn results in autophosphorylation of the RTK
(Schlessinger 2000; Neer 1995). Autophosphorylation of tyrosine resi
dues in receptors creates binding sites for proteins containing Src-ho
mology-2 (SH2) and phosphotyrosine-binding (PTB) domains. These
interacting proteins may serve as adaptors and/or show enzymatic activ
ities [e.g., ligand-regulated guanine nucleotide exchange factors (GEFs)
that regulate the function of the particular RTKJ. Alteration of either the
adaptor function or the enzymatic activity may affect the cascade of
protein-protein and protein-lipid interactions, phosphorylations, and
dephosphorylations, and the production of secondary messengers, that
lead ultimately to altered gene transcription and cellular function.
It has been observed that many RTKs are rapidly endocytosed on li
gand activation, after which they traffic through the endomembrane net
work, an elaborate system of interconnecting tubules and vesicles that
mediate the transport of fluid and selected membrane proteins. The sug
gestion that these endocytic membranes have a role in cell signaling has
been an open question.
Cell surface receptors have also served as model systems for the study
of general mechanisms and pathways of endocytic transport. Early stud
ies of the relationship between signaling and endocytosis relied mostly
on the use of subcellular fractionation of membrane compartments and
on the analysis of receptors harboring mutations. More recently, experi
mental approaches to interfere with specific mechanisms of membrane
transport have been developed, allowing a detailed study of specific sig
naling and membrane transport events in living cells.
In this review, we highlight the close ties between endocytosis and
signaling by discussing important observations from early studies along
with selected recent studies that have provocative implications.
2
Endocytosis of Signaling Receptor
The most comprehensive studies of RTK endocytosis have been carried
out with the epidermal growth factor (EGF) receptor as a model system
(Fig. 1). Cellular stimulation with EGF results in rapid clustering of
EGF receptor complexes in clathrin-coated pits and the subsequent in
ternalization into clathrin-coated endocytic vesicles (Gorden et al. 1978;
Receptor Tyrosine Kinase Signaling and Trafficking-Paradigms Revisited 3
Recydm,
Endosome
olgi
ompl .
Fig. 1. Endocytic pathway: This figure shows the progression of an endocytic vesicle
from internalization to maturation at the lysosome. RTKs are used to illustrate the
possible fates of an endocytosed receptor complex. The various Rab GTPases
thought to be involved in this pathway are also indicated. Rab22 may playa role in
membrane sorting in the early endosome (Mesa et al. 2001)
Hanover et al. 1984; Carpentier et al. 1982). The internalization of EGF
receptors can be effectively blocked by dominant-negative mutants of
several regulatory proteins. Examples include dynamin, a cytoplasmic
GTPase that is necessary for the fission of coated vesicles from the plas
ma membrane, and Rab5, a cytoplasmic GTPase that is necessary for en
dosome fusion (Barbieri et al. 2000; Damke et al. 1994; Carbone et al.
1997). Under physiological conditions, coated pits are the main routes
for internalization of growth factor receptors. However, these receptors
have also been shown to internalize by clathrin-independent processes
that resemble micropinocytosis, particularly in cells over expressing the
receptor (Haigler et al. 1979; Hopkins et al. 1985).
