Abstract
Salmonella invades mammalian cells by inducing membrane ruffling and macropinocytosis through actin remodelling. Because phosphoinositides are central to actin assembly, we have studied the dynamics of phosphatidylinositol-4,5-bisphosphate (PtdIns(4,5)P2) in HeLa cells during invasion by Salmonella typhimurium. Here we show that the outermost parts of the ruffles induced by invasion show a modest enrichment in PtdIns(4,5)P2, but that PtdIns(4,5)P2 is virtually absent from the invaginating regions. Rapid disappearance of PtdIns(4,5)P2 requires the expression of the Salmonella phosphatase SigD (also known as SopB). Deletion of SigD markedly delays fission of the invaginating membranes, indicating that elimination of PtdIns(4,5)P2 may be required for rapid formation of Salmonella-containing vacuoles. Heterologous expression of SigD is sufficient to promote the disappearance of PtdIns(4,5)P2, to reduce the rigidity of the membrane skeleton, and to induce plasmalemmal invagination and fission. Hydrolysis of PtdIns(4,5)P2 may be a common and essential feature of membrane fission during several internalization processes including invasion, phagocytosis and possibly endocytosis.
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Acknowledgements
This work was supported by the Arthritis Society of Canada and the Canadian Institutes for Health Research. M.T. is the recipient of a Hospital for Sick Children Restracomp Fellowship. O.V.V. is the recipient of a postdoctoral fellowship from PRAXIS XX1. B.B.F and S.G. are recipients of Canadian Institutes for Health Research (CIHR) Distinguished Scientist Awards. S.G. is the current holder of the Pitblado Chair in Cell Biology at The Hospital for Sick Children and is cross-appointed to the Department of Biochemistry, University of Toronto. C.M.Y. is the holder of a Canada Research Chair.
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Movie 1
Live imaging of HeLa cells during invasion by wildtype Salmonella enterica (serovar Typhimurium). (MOV 2714 kb)
Movie 2
Live imaging of HeLa cells during invasion by mutant Salmonella lacking SopB/SigD. (MOV 2988 kb)
Movie 3
Live imaging of HeLa cells during invasion by mutant Salmonella lacking SopE and SopE2 (MOV 2899 kb)
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Terebiznik, M., Vieira, O., Marcus, S. et al. Elimination of host cell PtdIns(4,5)P2 by bacterial SigD promotes membrane fission during invasion by Salmonella. Nat Cell Biol 4, 766–773 (2002). https://doi.org/10.1038/ncb854
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DOI: https://doi.org/10.1038/ncb854
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