Table Of ContentProgress in Molecular and Subcellular Biology
Series Editors: W.E.G.Muller (Managing Editor),Ph. Ieanteur, 28
I. Kostovic,Y.Kuchino,A.Macieira-Coelho,R.E.Rhoads
Springer-Verlag Berlin Heidelberg GmbH
A.-P. Arrigo· W.E.G. Muller (Eds.)
Small Stress
Proteins
With 47 Figures
Springer
Professor Dr. ANDRE-PATRICK ARRIGO
Laboratoire Stress Oxydant
Chaperons et Apoptose
Centre de Genetique Moleculaire et Cellulaire
CNRS UMR-5534
Universite Claude Bernard Lyon-I
43, Bd. du 11 Novembre
69622 Villeurbanne, France
W.E.G. MOLLER
Institut fUr Physiologische Chemie
Abteilung fur Angewandte Molekularbiologie
Johannes Gutenberg-Universitiit
Duesbergweg 6
55099 Mainz, Germany
ISSN 0079-6484
ISBN 978-3-642-62708-8
Library of Congress Cataloging-in-Publication Data
Small stress proteins / Andn!-Patrick Arrigo, Werner E.G. Mtiller (eds.).
p. cm. - (Progress in molecular and subcellular biology ; 28)
Includes bibliographical references and index.
ISBN 978-3-642-62708-8 ISBN 978-3-642-56348-5 (eBook)
DOI 10.1007/978-3-642-56348-5
1. Heat shock proteins. 1. Arrigo, Andn!-Patrick, 1947- II. Miil!er, Werner E.G. III. Series.
QP552.H43 563 2002 572'.6 - dc21
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Preface
Studies haveidentified important families ofproteins (denoted:heat shock or
stress proteins, Hsps) which display an enhanced expression in response to
heat shock or other physiological stresses. Besidesthe characterization of the
genes encoding Hsp and the mechanisms of their induction, recent studies
have concentrated on the function of these proteins. It was shown that the
expression of Hsp protects the cell against different types of aggressions.In
addition,Hsp can regulate essential biochemicalprocesses in unstressed cells.
For example, members of the Hsp60 and Hsp70 families act as ATP-binding
proteins allowing the folding of nascent or denatured proteins as well as the
assembly or disassemblyofproteincomplexes.These observationshaveled to
the discovery of the molecular chaperone concept (Ellis and Hemmingsen
1989).
Amongst the proteins whose expression is up-regulated by heat shock
or other types of stresses are the small stress proteins also denoted (sHsps,
sHsp or sHSP).Small stress proteins encompass a large numbers of related
proteins which are represented in virtually all organisms, including prokary
otes. These polypeptides share a structural domain, often referred to as the
a-crystallin domain, common to the lens protein alpha-crystallin (Ingolia
and Craig 1982;Wistow 1985).In addition to being increased in response to
several types ofstresses, the Hsp levelisalso upregulatedduring development
and correlates with the differentiation and oncogenic status of the cell. In
spite ofthe fact that sHsp can confer cellular protection againststresses,their
molecularfunction has remainedenigmaticforyears.Recently,majorfindings
have clarified this point. First, it has been shown that an ATP-independent
chaperone activity is associated to sHsp (Jacob and Buchner 1993).Asecond
important observation concerns the ability of sHsp to form large oligomers
which interact with misfolded polypeptides. These structures probably act
as reservoirs of folding intermediates that are subsequently presented to the
ATP-dependent chaperones (Hsp70 and other co-chaperones) (Ehrnsperger
et al. 1997; Lee et al. 1997). sHsp have therefore been called the "forgotten
chaperones".
In addition to their protein chaperone property,several otherobservations
have contributed to the renewed interest in sHsp.For example, human Hsp27
isexpressedin numerous cancer cellsand induces protection against avariety
of toxic chemicals used in chemotherapy. Hsp27 and aB-crystallin are also
expressed in several pathological situations, such as neurodegenerative dis-
VI Preface
eases.Moreover,amutation in aB-crystallin has been shown tobe responsible
foraspecial type ofmyopathy.Recentdata also suggest homeostaticfunctions
of sHsp at the levelof signal transduction,growth,differentiation and trans
formation processes. Anotherimportantcluewasthe recent finding thatsHsp
can protect against cellsuicide or"apoptosis"(Arrigo 1998,2000).
This book is the first one to be devoted only to small stress proteins. It
surveys the current knowledge concerning the expression and function of
sHsp in differentorganisms rangingform prokaryotesto human,except plant
sHsp which are not described here.Each chapter givesan overview of a spe
cificsubjectarea and includes currentresults from each author's laboratoryas
wellas a overviewof future research directions.This bookwillbe ofvalue to
researchers and graduate students interested in the fieldof cellstress biology,
rangingfrom the molecularto clinical level.
The book is organized in several parts describing different biological
systemsthatarecurrentlyusedtoanalyzethe expressionand function ofsHsp.
The first section of the book, which contains two chapters, deals with sHsp
genetic diversity. The first chapter provides an overview of the complex evo
lutionarydiversityofsHsps,particularlyprokaryoticsHspwhich arenowcon
sideredasancestors of eukaryotic sHsps.The second chapter deals with sHsp
diversity in a desert fish model. The second section of the book is related to
the function and expression of sHsp in different eukaryote models. It begins
with a chapter describing the chaperone activity of sHsp.Then, sHsp from C.
elegans,Drosophila and different mammaliansystems are presented.The third
section ofthe bookcontains three chapterswhich describe pathologicalstates
associated with sHsp expression and gene therapyapproaches aimed at mod
ulating sHsp levelof expression.
Wewouldliketoexpressour gratitudetoallauthorswhocontributedtothis
book for their cooperation and patience and to Springer for its support and
strong interest in publishingthis book.
In this book, smallstressproteins (cognate orstressinducedandabbreviated
sHsp,sHsps or sHSPdepending the authors) are defined as those proteinspos
sessing the so-called a-crystallinprotein domain. The name usedfor individual
sHsp is HspXX where xx corresponds to the two first digits ofthe apparent
molecular weight, excluding aA and aB-crystallins which are called by their
own name.
Pers-Iussy,France A.-P.ARRIGO,W.E.GMOLLER
August 2001
References
ArrigoAP(1998)Small stressproteins:chaperonesthat act as regulators of intracellularredox
stateand programmedcelldeath. BioIChern379:19-26
ArrigoAP(2000)sHsp asnovelregulatorsofprogrammedcelldeathandtumorigenicity.Pathol
BioI(Paris)48:280-288
Preface VII
EhrnspergerM,Graber S,Gaestel M,Buchner J(1997)Binding ofnon-native proteinto Hsp25
during heat shockcreates a reservoir of folding intermediates for reactivation. EMBOJ 16:
221-229
EllisRJ,VanderViesSM,Hemmingsen (1989)The molecular chaperone concept.Biochem Soc
Symp55:145-153
Ingolia TD,CraigE(1982)Four small Drosophilaheat shockproteins are related to each other
andtomammalianalpha-crystallin.Proc Nat!AcadSciUSA79:2360-2364
JakobU,GaestelM,EngelK,BuchnerJ(1993)Smallheat shockproteinsare molecular chaper
ones.JBioiChern268:1517-1520
LeeGJ, Roseman AM, Saibil HR,Vierling E (1997) A small heat shock protein stably binds
heat-denaturedmodelsubstratesand can maintainasubstrateinafolding-competentstate.
EMBOJ16:659-671
Wistow G (1985) Domain structure and evolution in alpha-crystallins and small heat shock
proteins.FEBSLett 181:1-6
Contents
Evolution and DiversityofProkaryoticSmallHeat ShockProteins
G.Kapp, J.A.M.Leunissen,andw.w.de long
1 Introduction . 1
2 sHsps in Prokaryotes . 2
3 Are sHsps Dispensable in Some Pathogenic Bacteria? . 3
4 PhylogeneticAnalysis ofProkaryotic sHsps . 7
5 Lateral Transfer or Convergent Evolution ofProkaryotic
sHsp Genes . 13
6 SecondaryStructure Prediction ofthe a-Crystallin
Domain . 14
7 Alignment and SecondaryStructure Prediction
ofthe N-Terminal Domain . 14
8 Summaryand Concluding Remarks . 15
References . 16
DiscoveryofTwoDistinctSmallHeatShockProtein (HSP) Families
in the DesertFish Poeciliopsis
C.E.Norris and L.E.Hightower
1 Introduction ... ... .... .. ... .... . .... ... .. ... ... .... .. 19
2 Variation in Small HSPsAmong Desert Species
ofPoeciliopsis ..... ................................... 20
2.1 Results ofSurveyAmong Species 20
2.2 HSP Isoforms as PrimaryGene Products 21
2.3 Conclusions .. ... ... .. ... .... ... ... ... .. ... ... .... ... 22
3 Variation in Small HSPsWithin Poeciliopsis Species .. ... ... .. 24
3.1 Results of Within-Species Survey. .. . .... ... .. ... .. .... ... 24
3.2 Conclusions. .. ... ... ... ... .... .. .... .. ... .. ... ... ... 25
4 DiscoveryofTwoClasses of Small HSPs in a Single Taxon .... 26
4.1 Differential Synthesis . ... ... ... ... .. .... .. ... .. ... ... .. 26
4.2 Phosphorylation State ofthe Small HSPs 28
4.3 SequenceAnalysis 28
4.4 EvolutionaryAnalysis .... ... .. .. .. ... ... ... .. ... .... ... 31
4.5 Conclusions. .. .. .... ... .. ..... .. .. .... .. ... ... .... .. 33
References ... ... ... ... ... ... .... . .... ... .. .. ... ... ... 34
x Contents
Chaperone Functionof sHsps
M.Haslbeck and J. Buchner
1 Introduction . 37
2 GeneralAspect of Chaperone Function . 37
2.1 Chaperones . 37
2.2 Analysis of Chaperone Function . 39
3 The Chaperone Properties of sHsps . 41
3.1 In Vivo Functions ofsHsps . 41
3.2 sHsps Bind Non-Native Protein . 44
3.3 Complex Formation and Substrate Range . 45
3.4 The Substrate BindingSite . 46
3.5 Mechanism of Substrate Binding . 47
3.6 Regulation of Functional Properties . 48
3.7 sHsps in the Context of the Cellular Chaperone Machinery . 49
4 Conclusions . 50
References . 52
The Small HeatShockProteinsoftheNematode Caenorhabditis
elegans:Structure,Regulationand Biology
E.Peter M.Candido
1 Introduction . 61
2 The Small Heat Shock Protein Family of C.elegans . 61
3 The 16-kDa Stress Proteins (HspI6s) . 63
3.1 Hsp16 Gene Organization . 63
3.2 Hsp16 Regulation . 65
3.3 Biochemical Properties of Hsp16s . 68
3.4 Usesof Hsp16 Promoters in C.elegans Research . 69
4 The 12-kDa Small Heat Shock Proteins (HsplZs) . 70
5 SIP-l ... .... .•...... ... ..................... .. ..... . 73
6 Hsp17.5 . 73
7 Hsp25 . 73
8 Hsp43 . 75
9 Conclusions and Prospects . 75
References . 76
Drosophila SmallHeat ShockProteins:CellandOrganelle-Specific
Chaperones?
S.Michaud,G.Morrow,J. Marchand,and R.M.Tanguay
1 Introduction ......................................... 79
2 Members of the Small Heat Shock Proteins Family 80
3 Transcriptional Regulation 80
3.1 Organisation ofChromatin 80
3.2 Stress-Induced Activation by the Heat Shock Factor 82
Contents XI
3.3 Developmental Transcription of shspGenes . 83
3.3.1 Regulation Cascade Induced by Ecdysone . 83
3.3.2 Tissue- and Cell-SpecificEnhancers . 85
4 Intracellular Localisation-Analysis of Targeting Signals . 85
5 Biochemical Properties and Post-Translational
Modifications . 86
6 Stress-Induced Expression of sHsp . 87
6.1 Cell-SpecificResponse of sHsp . 88
6.2 Functions ofsHsp Under Stress Conditions . 88
7 Developmental Expression of sHsp . 90
7.1 Stage,Tissue,and CellSpecificity . 90
7.1.1 Embryogenesis . 90
7.1.2 Germ Line . 92
7.1.3 Ageing . 92
7.2 Functions of sHsp During Normal Development . 93
7.2.1 Interactionwith the SUMO-Conjugating Enzyme Ubc9 . 94
7.2.2 Modulation of SpecificBiologicalActivity . 94
8 Conclusion . 96
References . 97
The DevelopmentalExpression of Small HSP
S.M.Davidson, M.-T.Loones, O.Duverger, and M.Morange
1 Introduction ......................................... 103
2 SmallHSPin Extraembryonic Development ................ 104
3 SmallHSPin Ectodermal Lineages ....................... 104
3.1 Neural Crest CellDerivatives ............................ 104
3.2 Neurectoderm ....................................... 105
3.3 Expression in the Skin .. ............................... 106
4 Small HSPin Mesodermal Lineages 107
4.1 SmallHSPin Muscle Development 107
4.1.1 The Expression Pattern of sHSPin the Heart .... 107
4.1.2 The Expression Pattern of sHSPin Skeletal
and Smooth Muscle 109
4.1.3 The Subcellular Localization of sHSPin Muscle ............. 110
4.2 Small HSPin Other Mesodermal Lineages 110
4.2.1 Expression in Cartilageand Bones 110
4.2.2 Expression in the Derivatives of the Notochord III
4.2.3 Expression in the Kidney ............................... III
4.2.4 Expression in the Adrenal Glands III
4.2.5 Expression in the Gonads 112
4.2.6 Expression in the Blood Cells 112
5 Small HSPin Endodermal Lineages 112
5.1 Expression in the Upper Digestive Tract ................... 112
5.2 Expression in the StomachAnd Intestine 112
