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Elimination of host cell PtdIns(4,5)P2 by bacterial SigD promotes membrane fission during invasion by Salmonella

Nature Cell Biology volume 4, pages 766–773 (2002)Cite this article

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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Figure 1: Effect of Salmonella on PtdIns(4,5)P2 in HeLa cells.
Figure 2: Use of FM4-64 to visualize the plasmalemma during Salmonella invasion.
Figure 3: Effect of Salmonella deficient in either SigD or SopE and SopE2 on PtdIns(4,5)P2.
Figure 4: Role of SigD in the elimination of PtdIns(4,5)P2.
Figure 5: Transfection of SigD into mammalian cells.
Figure 6: Detection of PtdIns(4,5)P2 using monoclonal antibodies.
Figure 7: SigD alters membrane elasticity and induces vacuole formation.
Figure 8: Effect of SigD on Salmonella vacuole sealing.

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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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Authors and Affiliations

  1. Cell Biology Program, Hospital for Sick Children, Toronto, M5G 1X8, Ontario, Canada

    Mauricio R. Terebiznik, Otilia V. Vieira, William S. Trimble & Sergio Grinstein

  2. Biotechnology Laboratory, University of British Columbia, Vancouver, V6T 1Z3, British Columbia, Canada

    Sandra L. Marcus & B. Brett Finlay

  3. Department of Biochemistry, University of Toronto, Toronto, M5S 3G9, Ontario, Canada

    Andrea Slade & Christopher M. Yip

  4. Department of Chemical Engineering, University of Toronto, Toronto, M5S 3G9, Ontario, Canada

    Christopher M. Yip

  5. Department of Molecular Pharmacology, Stanford University, Stanford, California, USA

    Tobias Meyer

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Supplementary information

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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