The Chlamydia trachomatis IncA protein is required for homotypic vesicle fusion.
Chlamydiae replicate within an intracellular vacuole, termed an inclusion, that is non-fusogenic with vesicles of the endosomal or lysosomal compartments. Instead, the inclusion appears to intersect an exocytic pathway from which chlamydiae intercept sphingomyelin en route from the Golgi apparatus to the plasma membrane. Chlamydial protein synthesis is required to establish this interaction. In an effort to identify those chlamydial proteins controlling vesicle fusion, we have prepared polyclonal antibodies against several Chlamydia trachomatis inclusion membrane proteins. Microinjection of polyclonal antibodies against three C. trachomatis inclusion membrane proteins, IncA, F and G, into the cytosol of cells infected with C. trachomatis demonstrates reactivity with antigens on the cytoplasmic face of the inclusion membrane, without apparent inhibition of chlamydial multiplication. Microinjection of antibodies against the C. trachomatis IncA protein, however, results in the development of an aberrant multilobed inclusion structure remarkably similar to that of C. psittaci GPIC. These results suggest that the C. trachomatis IncA protein is involved in homotypic vesicle fusion and/or septation of the inclusion membrane that is believed to accompany bacterial cell division in C. psittaci. This proposal is corroborated by the expression of C. trachomatis and C. psittaci IncA in a yeast two-hybrid system to demonstrate C. trachomatis, but not C. psittaci, IncA interactions. Despite the inhibition of homotypic fusion of C. trachomatis inclusions, fusion of sphingomyelin-containing vesicles with the inclusion was not suppressed.
Hackstadt
T
T
Laboratory of Intracellular Parasites, NIAID, NIH, Hamilton, MT 59840, USA. ted_hackstadt@nih.gov
Scidmore-Carlson
M A
MA
Shaw
E I
EI
Fischer
E R
ER
D000907:Antibodies, Bacterial / Q000276:immunology; D001426:Bacterial Proteins; D002692:Chlamydia trachomatis / Q000235:genetics / Q000276:immunology / Q000502:physiology* / Q000648:ultrastructure; D006367:HeLa Cells; D006801:Humans; D002479:Inclusion Bodies / Q000382:microbiology* / Q000502:physiology / Q000648:ultrastructure; D008561:Membrane Fusion / Q000502:physiology*; D008845:Microinjections; D008854:Microscopy, Electron; D010750:Phosphoproteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D013109:Sphingomyelins / Q000378:metabolism; D013997:Time Factors; D022161:Transport Vesicles / Q000378:metabolism*; D020798:Two-Hybrid System Techniques
D000907:Antibodies, Bacterial; D001426:Bacterial Proteins; C094163:IncA protein, bacteria; D010750:Phosphoproteins; D013109:Sphingomyelins