Chlamydial secretions systems

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Table of contents

TABLE OF CONTENTS
TABLE OF FIGURES
ABBREVIATIONS
ABSTRACT
RESUME (UNE PAGE)
RESUME (VERSION LONGUE)
INTRODUCTION
1. Chlamydiae
1.1 Phylogeny
2. Human pathogens
2.1 C. trachomatis
2.1.1 Trachoma, disease and history
2.1.2 Urogenital tract infections
2.1.3 Lymphogranuloma venereum
2.1.4 Diagnosis and treatment
2.2 C. pneumoniae
3. Biphasic developmental cycle
3.1 Entry to Exit
3.1.1 Attachment and Entry
3.1.2 Building the inclusion and survival
3.2 Characteristics of EBs and RBs
3.2.1 Ultrastructure of the chlamydial surface
3.2.2 Transcriptional regulation
3.3 Persistence
4. Host-pathogen interactions
4.1 Chlamydial secretions systems
4.2 Recruitment and uptake of host organelles and lipids
4.2.1 Recruitment of host lipids
4.2.2 Interaction with host organelles
4.3 Interface of interaction: the inclusion membrane
4.4 The host defence and chlamydial immune evasion
5. Chlamydial metabolism
5.1 Energy parasite hypothesis
5.2 Metabolic pathways in the post-genomic era
5.2.1 Substrate level phosphorylation
5.2.2 Oxidative phosphorylation
5.2.3 Other metabolic pathways
5.3 Glycogen
5.3.1 Chlamydial glycogen metabolism
5.3.2 Eukaryotic glycogen metabolism
MATERIAL AND METHODS
Cells and bacteria
Electron microscopy and periodic acid-thiocarbohydrazide-silver proteinate reaction
(PATAg)
Reinfection assay and flow cytometry
Quantitative Reverse Transcription PCR and Reverse Transcription PCR
Transfection
Construction of recombinant plasmids
Immunofluorescence and PAS staining
Quantification of glycogen with CellProfiler
Western Blot and antibodies
Zymogram
Heterologous secretion assay in Shigella flexneri
Transformation of C. trachomatis L2
Glucose uptake assay
RESULTS
1. Glycogen detection in Chlamydia trachomatis inclusion
2. Glucose is essential for Chlamydia trachomatis infection
3. Intraluminal glycogen is not derived from bacterial lysis
4. Kinetics of glycogen accumulation
5. Mechanisms of glycogen accumulation in inclusion lumen
5.1 Hypotheses
5.2 Vesicular import of host glycogen
5.2.1 Part of luminal glycogen is translocated in bulk from the host cytoplasm
5.2.2 Import of host glycogen and glycogen enzymes is autophagosome independent
5.3 Import of host glucose derivative
5.3.1 UDP-glucose is the host sugar transported into the inclusion lumen
5.3.2 UDP-glucose is a substrate for chlamydial GlgA
5.3.3 Identification of UDP-Glc transporter at inclusion membrane
6. Chlamydial glycogen enzymes are secreted for de novo glycogen synthesis
6.1 Heterologous test of secretion in Shigella flexneri
6.2 GlgX is present in the inclusion lumen
6.3 Overexpression of glycogen enzymes in C. trachomatis
6.4 Ectopically expressed GlgA compensates for the plasmid-less deficiency in glycogen accumulation
7. Chlamydia import Glc6P, but not Glc1P nor Glc
DISCUSSION
BIBLIOGRAPHY