Institut de Biologie Moléculaire et Cellulaire CNRS UNIVERSITE DE STRASBOURG

Stefanie Limmer

Description

Niveau: Supérieur, Doctorat, Bac+8
Institut de Biologie Moléculaire et Cellulaire CNRS – UNIVERSITE DE STRASBOURG THESE DE DOCTORAT Discipline: Sciences du vivant Aspects Moléculaires et Cellulaires de la Biologie En vue d'obtenir le grade de Docteur de l'Université de Strasbourg Stefanie LIMMER Etude des relations hôte-pathogène dans des modèles d'infection intestinales de Drosophila melanogaster Soutenue le 19 Octobre 2010 devant la commission d'examen: Prof. Arturo ZYCHLINSKY (Rapporteur Externe), Max Planck Institut für Infektionsbiologie, Berlin Dr. Jonathan EWBANK (Rapporteur Externe), U631 de l'INSERM, Centre d'Immunologie, Marseille Prof. Philippe GEORGEL (Examinateur), Laboratoire d'Immunogénétique Moléculaire, Strasbourg Prof. Jules HOFFMANN (Examinateur), UPR9022 du CNRS, Strasbourg Prof. Christian KLÄMBT (Examinateur), Institut für Neurobiologie, Münster Dr. Dominique FERRANDON (Directeur de thèse), UPR9022 du CNRS, Strasbourg

• spz spätzle


• diap2 drosophila


• c4-hsl butanoyl-homoserine lactone


• modèles d'infection intestinales de drosophila melanogaster


• mitogen activated protein


• drosophila melanogaster


• spe spätzle-processing-enzyme


• kinase


• enzyme pqs

Sujets

• Georgel
• Hoffmann
• Planck
• Université de Strasbourg
• Strasbourg
• Carpet sweeper

Informations

• Publié par: profil-zyak-2012
• Publié le: 01 octobre 2010
• Nombre de lectures: 19
• Langue: English
• Poids de l'ouvrage: 9 Mo

Extrait

Institut de Biologie Moléculaire et Cellulaire
CNRS – UNIVERSITE DE STRASBOURG




THESE DE DOCTORAT
Discipline: Sciences du vivant
Aspects Moléculaires et Cellulaires de la Biologie

En vue d’obtenir le grade de
Docteur de l’Université de Strasbourg



Stefanie LIMMER



Etude des relations hôte-pathogène dans des modèles
d’infection intestinales de Drosophila melanogaster





Soutenue le 19 Octobre 2010 devant la commission d’examen:

Prof. Arturo ZYCHLINSKY (Rapporteur Externe), Max Planck Institut für Infektionsbiologie, Berlin
Dr. Jonathan EWBANK (Rapporteur Externe), U631 de l’INSERM, Centre d'Immunologie, Marseille
Prof. Philippe GEORGEL (Examinateur), Laboratoire d’Immunogénétique Moléculaire, Strasbourg
Prof. Jules HOFFMANN (Examinateur), UPR9022 du CNRS, Strasbourg
Prof. Christian KLÄMBT (Examinateur), Institut für Neurobiologie, Münster
Dr. Dominique FERRANDON (Directeur de thèse), UPR9022 du CNRS, Strasbourg
Acknowledgment

First, I want to thank Prof. Jules Hoffmann and Prof. Jean-Marc Reichhart for having
made the laboratory one of the best in the field of immunology and a fruitful
environment for young researchers.

I also want to thank Prof. Arturo Zychlinsky, Dr. Jonathan Ewbank, Prof. Philippe
Georgel, Prof. Jules Hoffmann and Prof. Christian Klämbt for taking the time to read
and judge my work.

Dominique, over all the years you were always there to help and encourage me. You
also took the time for many, many fruitful discussions. I’m really happy that I had the
possibility to work in your group. I learned a lot during my time in Strasbourg. Thank
you.

Nadine, thank you for establishing the Serratia model and for introducing me into the
subject.

A big thanks to Cordu, Ioannis, Safia, Basti, Magda, Stan, Jessica, Ayyaz and Sunny
for their friendship and help. I’ll miss you all. Steffi, what should I say? Thank you! I
also want to thank all the other current and former members of the team and the rest
of the lab for all their help and support. Samantha: good luck for your thesis.

Last but not least, I want to thank my family and my boyfriend for their support and
encouragement over the years. Without you I would not have managed. Thank you. Abbreviations

3-oxo-C -HSL 3-oxo-dodecanoyl-homoserine lactone 12
AMP anti-microbial peptide
C -HSL butanoyl-homoserine lactone 4
CF cystic fibrosis
DAP diaminopimelic acid
DIAP2 Drosophila inhibitor of apoptosis 2
Dif Dorsal related immunity factor
Dome Domeless
Dscam Down syndrome cell adhesion molecule
DUOX dual oxidase
EB enteroblast
EC enterocyte
EMS ethyl methanesulfonate
Erk extracellular signal-regulated kinase
GlcNAc N-acetylglucosamine
GNBP gram-negative binding protein
GPCR G-protein coupled receptor
Gprk G-protein-coupled receptor kinase
hFAF1 human Fas associated factor 1
Hop Hopscotch
IAP inhibitor of apoptosis
IBM IAP-binding motif
IMD Immune deficiency
IP3 1,4,5-triphosphate
IRAK IL-1R associated kinase
ISC intestinal stem cell
Key Kenny
LPS lipopolysaccharide
LRR leucine-rich repeats
Lys lysine
MAMP microbe associated molecular pattern
MAPK mitogen-activated protein kinase ModSP modular serine protease
MurNAc N-acetylmuramic acid
N-acyl-HSL N-acylhomoserine lactone
NADPH nicotineamide adenine dinucleotide phosphate
ORF open reading frame
PGN peptidoglycan
PGRPs PGN-recognition proteins
PLCβ phospholipase C-β
PO phenoloxidase
PPAE Prophenoloxidase activating enzyme
PQS Pseudomonas Quinolone Signal
ProPO prophenoloxidase
PRRs pattern recognition receptors
Psh Persephone
Pvf PDGF- and VEGF-related factor
Pvr PDGF- and VEGF-receptor related
QS quorum sensing
ROS reactive oxygen species
ROS reactive oxygen species
SPE Spätzle-processing-enzyme
Spz Spätzle
Tep Thioester-containing protein
TLR Toll-like receptor
Tot Turandot
TPSS two partner secretion system
upd unpaired
WntD wnt inhibitor of Dorsal
βGRP β-glucan recognition proteins
Content 1
1 Introduction.......................................................................................................... 3
1.1 Overview ...................................................................................................... 4
1.2 Drosophila melanogaster ............................................................................. 5
Systemic response .............................................................................................. 6
Recognition of microbes .................................................................................. 6
Recognition of Gram(-) bacteria via DAP-type Peptidoglycan (PGN)........... 7
Recognition of Gram(+) bacteria via Lys-type Peptidoglycan (PGN).......... 10
Recognition of fungi.................................................................................... 11
Signal transduction ........................................................................................ 12
Activation of the Toll pathway..................................................................... 12
The Toll pathway........................................................................................ 13
Negative regulation of the Toll pathway ..................................................... 15
The IMD pathway ....................................................................................... 15
Negative regulation of the IMD pathway..................................................... 17
The JAK/STAT pathway ............................................................................. 21
Immune effectors ........................................................................................... 24
Antimicrobial peptides ................................................................................ 24
Tep proteins ............................................................................................... 24
Other effectors............................................................................................ 25
Local immune responses................................................................................... 25
Physical barrier and hostile environment in the midgut .............................. 26
AMP expression ......................................................................................... 27
ROS production.......................................................................................... 28
Cellular immune response................................................................................. 29
Phagocytosis.............................................................................................. 30
Encapsulation............................................................................................. 30
Coagulation ................................................................................................ 31
Melanization ............................................................................................... 32
Other immune functions of hemocyte......................................................... 32
1.3 Serratia marcescens .................................................................................. 34
The bacterium ............................................................................................ 34
S. marcescens infection in Drosophila ....................................................... 36
1.4 Pseudomonas aeruginosa.......................................................................... 37
The bacterium ............................................................................................ 37
Content 2
P. aeruginosa infections in Drosophila ....................................................... 41
1.5 Aim of this work.......................................................................................... 41
2 Serratia marcescens infections ......................................................................... 43
2.1 Genome-Wide RNAi screen identifies genes involved in intestinal
pathogenic bacterial infection................................................................................ 44
Introduction........................................................................................................ 44
Additional results and discussion ...................................................................... 47
Validation of candidate genes .................................................................... 47
JAK/STAT pathway and compensatory proliferation .................................. 52
2.2 Six hour-long regeneration of the Drosophila melanogaster midgut following
its partial degradation by ingested Serratia marcescens....................................... 57
Introduction........................................................................................................ 57
Discussion ................................................