ADP-ribosylation by exoenzyme T of Pseudomonas aeruginosa induces an irreversible effect on the host cell cytoskeleton in vivo.
Pseudomonas aeruginosa utilises a type III secretion system (TTSS) to introduce exoenzyme S and exoenzyme T into host cells to subvert host cell signalling and thereby promote infection. In this study, we have employed the heterologous TTSS of Yersinia to deliver different mutants of ExoT into HeLa cells. Wild-type ExoT and ExoT variants expressing either GAP (GTPase activating protein) or ADP-ribosyltransferase activity mediated changes in cell morphology, which correlated to disruption of the actin microfilaments of the infected cells. ExoT expressing ADP-ribosylating activity gave an irreversible effect on HeLa cell morphology, while ExoT expressing only GAP activity displayed a reversible effect where the cells regained normal cell morphology after killing of the infecting bacteria. This shows that ExoT can modify and inactivate host cell proteins involved in maintaining the actin cytoskeleton in vivo by two independent mechanisms.
Sundin
Charlotta
C
Department of Medical Biosciences, Umeå University, Umeå, SE-90187 Sweden.
Hallberg
Bengt
B
Forsberg
Ake
A
D036002:ADP Ribose Transferases / Q000235:genetics / Q000378:metabolism*; D008841:Actin Cytoskeleton / Q000378:metabolism / Q000648:ultrastructure; D000199:Actins / Q000378:metabolism; D000246:Adenosine Diphosphate Ribose / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D048429:Cell Size; D003001:Cloning, Molecular; D003599:Cytoskeleton / Q000378:metabolism* / Q000648:ultrastructure; D020690:GTPase-Activating Proteins; D015870:Gene Expression; D005798:Genes, Bacterial; D006367:HeLa Cells; D006801:Humans; D008853:Microscopy; D008856:Microscopy, Fluorescence; D009154:Mutation; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D011550:Pseudomonas aeruginosa / Q000201:enzymology* / Q000472:pathogenicity; D011994:Recombinant Proteins / Q000378:metabolism; D015398:Signal Transduction; D015011:Yersinia pseudotuberculosis / Q000235:genetics / Q000378:metabolism
D000199:Actins; D001426:Bacterial Proteins; C477521:ExoT protein, Pseudomonas aeruginosa; D020690:GTPase-Activating Proteins; D011994:Recombinant Proteins; D000246:Adenosine Diphosphate Ribose; D036002:ADP Ribose Transferases