Thèse de Doctorat de l'Université Louis Pasteur Strasbourg I

Thèse de Doctorat de l’Université Louis Pasteur Strasbourg I
Discipline: Sciences du vivant


Présentée et soutenue publiquement par

Anne PACQUELET

Pour obtenir le grade de
Docteur de l’Université Louis Pasteur Strasbourg I


Sujet de thèse:

Régulation de l’adhérence médiée par
la DE-cadhérine au cours de la migration des
cellules de bordure chez la Drosophile




Soutenue le 02 juillet 2004 devant le jury composé de:

Mr Jean-Marc Reichhart Professeur, Strasbourg Directeur de thèse
Mme Pernille Rørth Directeur de recherche, Heidelberg Codirectrice de thèse
Mr Julien Royet Professeur, Strasbourg Rapporteur interne
Mr Stephen Cohen Directeur de recherche, Heidelberg Rapporteur externe
Mr François Schweisguth Directeur de recherche, Paris Rapporteur externe
Thèse de Doctorat de l’Université Louis Pasteur Strasbourg I
Disciplin: Biology


Defended by

Anne PACQUELET

To obtain the degree of
Docteur de l’Université Louis Pasteur Strasbourg I


Title:

Regulation of Drosophila E-cadherin mediated
adhesion during border cell migration






ndDefended on the 2 of July 2004 with the following jury:

Mr Jean-Marc Reichhart Professeur, Strasbourg Directeur de thèse
Mme Pernille Rørth Directeur de recherche, Heidelberg Codirectrice de thèse
Mr Julien Royet Professeur, Strasbourg Rapporteur interne
Mr Stephen Cohen Directeur de recherche, Heidelberg Rapporteur externe
Mr François Schweisguth Directeur de recherche, Paris Rapporteur externe
1
ACKNOWLEDGMENTS


Many people have made this work possible, and I am glad to acknowledge them here.

First of all, I would like to thank Pernille Rørth for giving me the chance to work in her lab
and for all her advices, enthusiasm and support during these years.

I would also like to thank Jean-Marc Reichhart for kindly accepting to be my supervisor at the
Université Louis Pasteur and being always available when needed. Thanks to Damian
Brunner and Steve Cohen who took the time to participate in my thesis committee at EMBL.
Thanks to Steve Cohen, Julien Royet and François Schweisguth for accepting to read my
thesis and be in my defence committee.


Thanks to all the people in the lab – past and current members: Andreea, Carlos, Gaspar,
Gemma, Hsin-Ho, Juliette, Kalman, Lodovica, Luis, Oguz, Peter, Simone and Tudor – for
tossing my flies on Sundays or taking care of my eppendorfs when I had to be at home, for all
our scientific and political discussions and for the great time I had in the lab. A special thank
to Li Lin who helped with the quantifications on the p120ctn project and cloned the DE-
cadherin-4YF mutant. I am also grateful to Ann Mari Voie who did all the injections to
generate transgenic flies.

Thanks to all the people in the fly labs and in EMBL who provided flies, reagents, advices or
friendly little talks in the corridors.


Merci à ma famille, en particulier à mes parents pour leur soutien pendant toutes ces années.

Et bien sûr un immense merci à Gwénaël, pour tout, tout simplement.
Merci aussi à Lucie, qui sans cesse nous rappelle à l’essentiel.
2
TABLE OF CONTENTS


SUMMARY................................................................................................................. 9
RÉSUMÉ (version courte) ..................................................................................... 10
RÉSUMÉ (version longue) ..................................................................................... 11
LIST OF ABBREVIATIONS ..................................................................................... 20

1 INTRODUCTION 23
1.1 Cell migration............................................................................................. 24
1.1.1 Importance...................................................................................................................24
1.1.2 Swimming and crawling............................................................................................ 24
1.1.3 Molecular mechanisms involved in cell crawling................................................... 25
1.1.3.1 Actin polymerization ....................................................................................26
1.1.3.2 Generation of traction forces.......................................................................29
1.1.3.3 Adhesion......................................................................................................29
1.2 Cadherin-mediated cell-cell adhesion...................................................... 32
1.2.1 The Cadherin superfamily......................................................................................... 32
1.2.1.1 A common feature: the cadherin repeat (CR) .............................................33
1.2.1.2 Classic cadherin ..........................................................................................34
1.2.1.3 Desmosomals cadherins.............................................................................36
1.2.1.4 Fat-like cadherins and seven-transmembrane cadherins...........................37
1.2.1.5 Protocadherins ............................................................................................37
1.2.2 Adhesion mechanisms.............................................................................................. 37
1.2.3 Role of classic cadherins during development ...................................................... 38
1.2.4 Molecules associated with classic cadherins at junctions ................................... 41
1.2.4.1 b-catenin and a-catenin: linking cadherin to actin......................................41
1.2.4.2 Other cytosolic proteins associated with cadherins ....................................43
1.2.4.3 Transmembrane proteins associated with cadherins..................................44
1.2.5 Regulation of cadherin mediated adhesion ............................................................ 44
1.2.5.1 Modulation of the link to actin filaments ......................................................45
1.2.5.2 Endocytosis .................................................................................................46 3
1.2.5.3 Regulation of lateral clustering....................................................................47
1.2.5.4 Regulation by p120 catenin.........................................................................47
1.2.5.5 Phosphorylation of cadherin and catenins ..................................................48
1.2.5.6 Small GTPases ...........................................................................................50
1.3 Border cell migration................................................................................. 52
1.3.1 Oogenesis and border cells...................................................................................... 52
1.3.2 Border cell specification ........................................................................................... 56
1.3.3 Guidance..................................................................................................................... 57
1.3.4 Extension formation and traction ............................................................................ 57
1.3.5 Adhesion during border cell migration ................................................................... 58
1.4 Aim of the project ...................................................................................... 60

2 RESULTS 61
2.1 p120ctn and juxtamembrane domain....................................................... 62
2.1.1 Strategy....................................................................................................................... 62
2.1.2 Controls ...................................................................................................................... 64
2.1.3 Neither interaction with p120ctn nor juxtamembrane domain are required
for DE-cadherin function during border cell migration ......................................... 68
2.1.4 Neither interaction with p120ctn nor juxtamembrane domain are required
for DE-cadherin function during oogenesis and development............................. 70
2.1.5 p120ctn itself does not seem to be required for Drosophila development ......... 74
2.2 Tyrosine phosphorylation......................................................................... 75
2.2.1 DE-cadherin tyrosine phosphorylation ................................................................... 75
2.2.1.1 Strategy .......................................................................................................75
2.2.1.2 DE-cadherin-4YF substitutes for endogenous DE-cadherin during
border cell migration ...................................................................................76
2.2.1.3 DE-cadherin-4YF substitutes for endogenous DE-cadherin during
oogenesis and development................................................................