Two coiled-coil domains of Chlamydia trachomatis IncA affect membrane fusion events during infection.
Chlamydia trachomatis replicates in a parasitophorous membrane-bound compartment called an inclusion. The inclusions corrupt host vesicle trafficking networks to avoid the degradative endolysosomal pathway but promote fusion with each other in order to sustain higher bacterial loads in a process known as homotypic fusion. The Chlamydia protein IncA (Inclusion protein A) appears to play central roles in both these processes as it participates to homotypic fusion and inhibits endocytic SNARE-mediated membrane fusion. How IncA selectively inhibits or activates membrane fusion remains poorly understood. In this study, we analyzed the spatial and molecular determinants of IncA's fusogenic and inhibitory functions. Using a cell-free membrane fusion assay, we found that inhibition of SNARE-mediated fusion requires IncA to be on the same membrane as the endocytic SNARE proteins. IncA displays two coiled-coil domains showing high homology with SNARE proteins. Domain swap and deletion experiments revealed that although both these domains are capable of independently inhibiting SNARE-mediated fusion, these two coiled-coil domains cooperate in mediating IncA multimerization and homotypic membrane interaction. Our results support the hypothesis that Chlamydia employs SNARE-like virulence factors that positively and negatively affect membrane fusion and promote infection.
Department of Microbiology and Immunology, Thomas Jefferson University, Philadelphia, Pennsylvania, United States of America.
Erik
E
Ronzone
Fabienne
F
Paumet
D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism*; D002690:Chlamydia Infections / Q000378:metabolism*; D002692:Chlamydia trachomatis / Q000378:metabolism*; D004705:Endocytosis; D006367:HeLa Cells; D006801:Humans; D008081:Liposomes / Q000378:metabolism; D008561:Membrane Fusion; D008565:Membrane Proteins / Q000737:chemistry* / Q000378:metabolism*; D055503:Protein Multimerization; D017434:Protein Structure, Tertiary; D050600:SNARE Proteins / Q000378:metabolism; D013329:Structure-Activity Relationship
D001426:Bacterial Proteins; C460868:IncA protein, Chlamydia trachomatis; D008081:Liposomes; D008565:Membrane Proteins; D050600:SNARE Proteins