Delineation and characterization of the actin nucleation and effector translocation activities of Salmonella SipC.
Salmonella type III secreted SipC possesses dual functions: translocation of effectors and actin modulation. The biological significance of SipC's actin nucleation activity in Salmonella-induced actin cytoskeleton rearrangements has not been studied. We report here the delineation of the actin nucleation activity from the effector translocation activity of SipC. Our data show that the central amino acid region (residues: 201-220) is essential for its actin nucleation activity and the C-terminal amino acid region (321-409) is required for translocation of effectors. A SipC nucleation-deficient mutant, which maintained its effector translocation activity, was obtained. This nucleation-deficient mutant had significantly reduced ability to induce actin cytoskeleton rearrangements, resulting in lower bacterial invasion into HeLa cells. Contrary to a previous report, we found that the purified recombinant wild-type SipC(199-409) protein is monomeric in solution by size exclusion chromatography coupled with multiangle laser light scattering assays (SEC-LS). Our data established that the actin nucleation activity of SipC plays a vital role in Salmonella-induced membrane ruffles and subsequent bacteria invasion.
Chang
JiHoon
J
Department of Biological Sciences, Purdue University, West Lafayette, IN 47907, USA.
Chen
Jue
J
Zhou
Daoguo
D
D000199:Actins / Q000737:chemistry / Q000378:metabolism*; D001426:Bacterial Proteins / Q000378:metabolism*; D003599:Cytoskeleton / Q000737:chemistry; D006367:HeLa Cells; D006801:Humans; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D012486:Salmonella typhimurium / Q000378:metabolism* / Q000472:pathogenicity; D014774:Virulence
D000199:Actins; D001426:Bacterial Proteins; D011993:Recombinant Fusion Proteins; C094578:Salmonella invasion protein C