The salmonella kinase SteC targets the MAP kinase MEK to regulate the host actin cytoskeleton

Charlotte Odendall; Nathalie Rolhion; Andreas Förster; John Poh; Douglas J. Lamont; Mei Liu; Paul S. Freemont; Andrew D. Catling; David W. Holden

doi:10.1016/j.chom.2012.09.011

The salmonella kinase SteC targets the MAP kinase MEK to regulate the host actin cytoskeleton

Charlotte Odendall
, Nathalie Rolhion
, Andreas Förster
, John Poh
, Douglas J. Lamont
, Mei Liu
, Paul S. Freemont
, Andrew D. Catling
, David W. Holden^*

^*Corresponding author for this work

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Research output: Contribution to journal › Article › peer-review

79 Citations (Scopus)

Abstract

After host cell entry, Salmonella replicate in membrane-bound compartments, which accumulate a dense meshwork of F-actin through the kinase activity of the Salmonella SPI-2 type III secretion effector SteC. We find that SteC promotes actin cytoskeleton reorganization by activating a signaling pathway involving the MAP kinases MEK and ERK, myosin light chain kinase (MLCK) and Myosin IIB. Specifically, SteC phosphorylates MEK directly on serine 200 (S200), a previously unstudied phosphorylation site. S200 phosphorylation is predicted to displace a negative regulatory helix causing autophosphorylation on the known MEK activatory residues, S218 and S222. In support of this, substitution of S200 with alanine prevented phosphorylation on S218 and S222, and phosphomimetic mutations of S200 stimulated phosphorylation of these residues. Both steC-null and kinase-deficient mutant strains displayed enhanced replication in infected cells, suggesting that SteC manipulates the actin cytoskeleton to restrain bacterial growth, thereby regulating virulence.

Original language	English
Pages (from-to)	657-668
Number of pages	12
Journal	Cell Host and Microbe
Volume	12
Issue number	5
DOIs	https://doi.org/10.1016/j.chom.2012.09.011
Publication status	Published - 15 Nov 2012

Bibliographical note

Funding Information:
We thank Emmanuelle Caron (Imperial College London, UK), Pascale Cossart (Pasteur Institute, France), Manuela Baccarini (University of Vienna, Austria), and Richard Marais (Institute of Cancer research, UK) for helpful discussions and members of the Holden laboratory for suggestions and critical reading of the manuscript. We thank Manuela Baccarini (University of Vienna, Austria) for MEFs KO for MEK1, Robert Adelstein (National Heart, Lung, and Blood Institute, Maryland, USA) for MEFs KO for Myosin IIB, and Cliona Boyle (Imperial College London, UK) for technical support. We thank Jonathan Kagan for his help and support. This work was supported by grants from the Medical Research Council and Wellcome Trust (UK) to D.W.H., P.F., and A.F, and from the European Molecular Biology Organization to N.R.

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title = "The salmonella kinase SteC targets the MAP kinase MEK to regulate the host actin cytoskeleton",

abstract = "After host cell entry, Salmonella replicate in membrane-bound compartments, which accumulate a dense meshwork of F-actin through the kinase activity of the Salmonella SPI-2 type III secretion effector SteC. We find that SteC promotes actin cytoskeleton reorganization by activating a signaling pathway involving the MAP kinases MEK and ERK, myosin light chain kinase (MLCK) and Myosin IIB. Specifically, SteC phosphorylates MEK directly on serine 200 (S200), a previously unstudied phosphorylation site. S200 phosphorylation is predicted to displace a negative regulatory helix causing autophosphorylation on the known MEK activatory residues, S218 and S222. In support of this, substitution of S200 with alanine prevented phosphorylation on S218 and S222, and phosphomimetic mutations of S200 stimulated phosphorylation of these residues. Both steC-null and kinase-deficient mutant strains displayed enhanced replication in infected cells, suggesting that SteC manipulates the actin cytoskeleton to restrain bacterial growth, thereby regulating virulence.",

author = "Charlotte Odendall and Nathalie Rolhion and Andreas F{\"o}rster and John Poh and Lamont, \{Douglas J.\} and Mei Liu and Freemont, \{Paul S.\} and Catling, \{Andrew D.\} and Holden, \{David W.\}",

note = "Funding Information: We thank Emmanuelle Caron (Imperial College London, UK), Pascale Cossart (Pasteur Institute, France), Manuela Baccarini (University of Vienna, Austria), and Richard Marais (Institute of Cancer research, UK) for helpful discussions and members of the Holden laboratory for suggestions and critical reading of the manuscript. We thank Manuela Baccarini (University of Vienna, Austria) for MEFs KO for MEK1, Robert Adelstein (National Heart, Lung, and Blood Institute, Maryland, USA) for MEFs KO for Myosin IIB, and Cliona Boyle (Imperial College London, UK) for technical support. We thank Jonathan Kagan for his help and support. This work was supported by grants from the Medical Research Council and Wellcome Trust (UK) to D.W.H., P.F., and A.F, and from the European Molecular Biology Organization to N.R. ",

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T1 - The salmonella kinase SteC targets the MAP kinase MEK to regulate the host actin cytoskeleton

AU - Odendall, Charlotte

AU - Rolhion, Nathalie

AU - Förster, Andreas

AU - Poh, John

AU - Lamont, Douglas J.

AU - Liu, Mei

AU - Freemont, Paul S.

AU - Catling, Andrew D.

AU - Holden, David W.

N1 - Funding Information: We thank Emmanuelle Caron (Imperial College London, UK), Pascale Cossart (Pasteur Institute, France), Manuela Baccarini (University of Vienna, Austria), and Richard Marais (Institute of Cancer research, UK) for helpful discussions and members of the Holden laboratory for suggestions and critical reading of the manuscript. We thank Manuela Baccarini (University of Vienna, Austria) for MEFs KO for MEK1, Robert Adelstein (National Heart, Lung, and Blood Institute, Maryland, USA) for MEFs KO for Myosin IIB, and Cliona Boyle (Imperial College London, UK) for technical support. We thank Jonathan Kagan for his help and support. This work was supported by grants from the Medical Research Council and Wellcome Trust (UK) to D.W.H., P.F., and A.F, and from the European Molecular Biology Organization to N.R.

PY - 2012/11/15

Y1 - 2012/11/15

N2 - After host cell entry, Salmonella replicate in membrane-bound compartments, which accumulate a dense meshwork of F-actin through the kinase activity of the Salmonella SPI-2 type III secretion effector SteC. We find that SteC promotes actin cytoskeleton reorganization by activating a signaling pathway involving the MAP kinases MEK and ERK, myosin light chain kinase (MLCK) and Myosin IIB. Specifically, SteC phosphorylates MEK directly on serine 200 (S200), a previously unstudied phosphorylation site. S200 phosphorylation is predicted to displace a negative regulatory helix causing autophosphorylation on the known MEK activatory residues, S218 and S222. In support of this, substitution of S200 with alanine prevented phosphorylation on S218 and S222, and phosphomimetic mutations of S200 stimulated phosphorylation of these residues. Both steC-null and kinase-deficient mutant strains displayed enhanced replication in infected cells, suggesting that SteC manipulates the actin cytoskeleton to restrain bacterial growth, thereby regulating virulence.

AB - After host cell entry, Salmonella replicate in membrane-bound compartments, which accumulate a dense meshwork of F-actin through the kinase activity of the Salmonella SPI-2 type III secretion effector SteC. We find that SteC promotes actin cytoskeleton reorganization by activating a signaling pathway involving the MAP kinases MEK and ERK, myosin light chain kinase (MLCK) and Myosin IIB. Specifically, SteC phosphorylates MEK directly on serine 200 (S200), a previously unstudied phosphorylation site. S200 phosphorylation is predicted to displace a negative regulatory helix causing autophosphorylation on the known MEK activatory residues, S218 and S222. In support of this, substitution of S200 with alanine prevented phosphorylation on S218 and S222, and phosphomimetic mutations of S200 stimulated phosphorylation of these residues. Both steC-null and kinase-deficient mutant strains displayed enhanced replication in infected cells, suggesting that SteC manipulates the actin cytoskeleton to restrain bacterial growth, thereby regulating virulence.

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JO - Cell Host and Microbe

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ER -

The salmonella kinase SteC targets the MAP kinase MEK to regulate the host actin cytoskeleton

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