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Table of contents
I.! Chlamydia trachomatis, a human pathogen
1.! Pathology & epidemiology: which diseases and where?
a.! Trachoma
b.! Urogenital infections
c.! Lymphogranuloma venereum
d.! Diagnosis and treatments
2.! A bit of history: once upon a time
3.! Chlamydia, a bacterium apart from others!
4.! The Chlamydia special: a biphasic developmental cycle
a.! Characteristics of EBs and RBs
b.! The very first step: the bacterial adhesion
c.! Knock-knock: The Chlamydia entry
d.! Home sweet home: Setting up the inclusion
e.! Multiplication and development: the reticulate bodies party
f.! The persistent form: a party forever?
g.! Parasitism of the host cell by Chlamydia
h.! The end of the show: a double way out ticket
5.! Chlamydia’s swiss knife: the secretion systems
a.! The T2SS: the general secretory pathway (GSP)
b.! The T5SS: the twin-arginine translocation (tat) pathway
c.! The T3SS: the chlamydial “injectisome”
6.! The challenge Chlamydia: from darkness to light
a.! A long time in the darkness
b.! … and then there was light: welcome to a new Chlamydia world.
II.! Interactions between Chlamydia and host membranes
1.! The first Chlamydia interaction: entering the cell
2.! Chlamydia fattens up: delivery of lipids to the inclusion
3.! A sociable guest: multiple interactions between Chlamydia and host organelles
a.! Chlamydia attracts most if not all the host organelles
b.! The Inc proteins: key components of the Chlamydia-host cell interaction
c.! Manipulation of the Rab proteins, key trafficking regulators
4.! Chlamydia trachomatis infection impairs cytokinesis
5.! The defensive answers of the host
6.! Chlamydia and ESCRT
III.! Biology of the ESCRT system
1.! Overview of a machinery with many complexes
2.! One system, many functions
a.! MVB biogenesis
b.! Abscission during cytokinesis
c.! Virus budding
d.! Exosomes secretion
e.! Autophagy
3.! Focus on Hrs
a.! From Hrs to the ESCRT-0 complex
b.! Dissection of Hrs: domains, interactions, regulations
c.! Hrs, a multi-functions protein
4.! Focus on Tsg101: a central protein for the ESCRT system
5.! ESCRT system, where else?
I.! Chlamydia strains
II.! Cell culture, transfection and chemicals
III.! Cloning procedures
IV.! Production and purification of recombinant protein
V.! Yeast-two-hybrid (Y2H) assays
VI.! Immunofluorescence and western blotting analysis
VII.! Quantification of bacterial entry
VIII.! Flow cytometry on infected cells and EBs
IX.! Immunoprecipitation (IP)
