title: Coiled-coil domains enhance the membrane association of Salmonella type III effectors.
abstract: Coiled-coil domains in eukaryotic and prokaryotic proteins contribute to diverse structural and regulatory functions. Here we have used in silico analysis to predict which proteins in the proteome of the enteric pathogen, Salmonella enterica serovar Typhimurium, harbour coiled-coil domains. We found that coiled-coil domains are especially prevalent in virulence-associated proteins, including type III effectors. Using SopB as a model coiled-coil domain type III effector, we have investigated the role of this motif in various aspects of effector function including chaperone binding, secretion and translocation, protein stability, localization and biological activity. Compared with wild-type SopB, SopB coiled-coil mutants were unstable, both inside bacteria and after translocation into host cells. In addition, the putative coiled-coil domain was required for the efficient membrane association of SopB in host cells. Since many other Salmonella effectors were predicted to contain coiled-coil domains, we also investigated the role of this motif in their intracellular targeting in mammalian cells. Mutation of the predicted coiled-coil domains in PipB2, SseJ and SopD2 also eliminated their membrane localization in mammalian cells. These findings suggest that coiled-coil domains represent a common membrane-targeting determinant for Salmonella type III effectors.
authors.affiliation: Laboratory of Intracellular Parasites, Rocky Mountain Laboratories, NIAID, NIH, Hamilton, Montana, USA. lknodler@niaid.nih.gov
authors.forename: Leigh A
authors.initials: LA
authors.lastname: Knodler
authors.forename: J Antonio
authors.initials: JA
authors.lastname: Ibarra
authors.forename: Ernesto
authors.initials: E
authors.lastname: Pérez-Rueda
authors.forename: Calvin K
authors.initials: CK
authors.lastname: Yip
authors.forename: Olivia
authors.initials: O
authors.lastname: Steele-Mortimer
mesh_terms: D002462:Cell Membrane / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D019295:Computational Biology; D018832:Molecular Chaperones / Q000378:metabolism; D016297:Mutagenesis, Site-Directed; D050505:Mutant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D020125:Mutation, Missense; D011485:Protein Binding; D011487:Protein Conformation; D055550:Protein Stability; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D012486:Salmonella typhimurium / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D037521:Virulence Factors / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*
chemical_list: D018832:Molecular Chaperones; D050505:Mutant Proteins; D037521:Virulence Factors