Abstract
Yersinia spp. cause gastroenteritis and the plague, representing historically devastating pathogens that are currently an important biodefense and antibiotic resistance concern. A critical virulence determinant is the Yersinia protein kinase A, or YpkA, a multidomain protein that disrupts the eukaryotic actin cytoskeleton. Here we solve the crystal structure of a YpkA-Rac1 complex and find that YpkA possesses a Rac1 binding domain that mimics host guanidine nucleotide dissociation inhibitors (GDIs) of the Rho GTPases. YpkA inhibits nucleotide exchange in Rac1 and RhoA, and mutations that disrupt the YpkA-GTPase interface abolish this activity in vitro and impair in vivo YpkA-induced cytoskeletal disruption. In cell culture experiments, the kinase and the GDI domains of YpkA act synergistically to promote cytoskeletal disruption, and a Y. pseudotuberculosis mutant lacking YpkA GDI activity shows attenuated virulence in a mouse infection assay. We conclude that virulence in Yersinia depends strongly upon mimicry of host GDI proteins by YpkA.
Publication types
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Research Support, N.I.H., Extramural
MeSH terms
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Amino Acid Sequence
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Animals
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Bacterial Proteins / chemistry
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Bacterial Proteins / genetics
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Bacterial Proteins / metabolism*
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Cell Line
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Crystallography
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Cytoskeleton / metabolism
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Guanine Nucleotide Dissociation Inhibitors / chemistry
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Guanine Nucleotide Dissociation Inhibitors / metabolism*
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Humans
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Intestinal Mucosa / metabolism
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Mice
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Models, Molecular
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Molecular Mimicry*
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Molecular Sequence Data
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Mutation
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Protein Serine-Threonine Kinases / chemistry
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Protein Serine-Threonine Kinases / genetics
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Protein Serine-Threonine Kinases / metabolism*
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Transfection
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Virulence Factors / chemistry
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Virulence Factors / genetics
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Virulence Factors / metabolism*
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Yersinia pseudotuberculosis / enzymology
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Yersinia pseudotuberculosis / metabolism
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Yersinia pseudotuberculosis / pathogenicity*
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cdc42 GTP-Binding Protein / chemistry
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cdc42 GTP-Binding Protein / metabolism
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rac1 GTP-Binding Protein / chemistry
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rac1 GTP-Binding Protein / metabolism*
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rho-Specific Guanine Nucleotide Dissociation Inhibitors
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rhoA GTP-Binding Protein / chemistry
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rhoA GTP-Binding Protein / metabolism
Substances
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Bacterial Proteins
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Guanine Nucleotide Dissociation Inhibitors
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Virulence Factors
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rho-Specific Guanine Nucleotide Dissociation Inhibitors
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ypkA protein, Yersinia
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Protein Serine-Threonine Kinases
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cdc42 GTP-Binding Protein
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rac1 GTP-Binding Protein
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rhoA GTP-Binding Protein