title: Two newly identified SipA domains (F1, F2) steer effector protein localization and contribute to Salmonella host cell manipulation.
abstract: Salmonella Typhimurium causes bacterial enterocolitis. The type III secretion system (TTSS)-1 is a key virulence determinant of S. Typhimurium mediating host cell invasion and acute enterocolitis. The TTSS-1 effector protein SipA is transported into host cells, accumulates in characteristic foci at the bacteria-host cell interface, manipulates signalling and affects virulence. Two functional domains of SipA have previously been characterized: The N-terminal SipA region (amino acids 1-105) mediates TTSS-1 transport and the C-terminal SipA 'actin-binding' domain (ABD; amino acids 446-685) manipulates F-actin assembly. Little is known about the central region of SipA. In a deletion analysis we found that the central SipA region harbours two distinct functional domains, F1 and F2. They are involved in SipA focus formation and host manipulation. The F1 domain (amino acids 170-271) drives SipA focus formation and domain F2 (amino acids 280-394) enhances this process by mediating SipA-SipA interactions. SipA variants lacking the F1-, the F2- or the actin binding domain were attenuated in virulence assays, namely host cell invasion and/or virulence in a mouse model for enterocolitis. Our results show that the newly identified SipA domains have distinct functions. Nevertheless, cooperation between the SipA domains F1, F2 and ABD is required to promote Salmonella virulence.
authors.affiliation: Institute of Microbiology, D-BIOL ETH Zürich, Wolfgang-Pauli Strasse 10, 8093 Zürich, Switzerland.
authors.forename: Markus C
authors.initials: MC
authors.lastname: Schlumberger
authors.forename: Rina
authors.initials: R
authors.lastname: Käppeli
authors.forename: Michael
authors.initials: M
authors.lastname: Wetter
authors.forename: Andreas J
authors.initials: AJ
authors.lastname: Müller
authors.forename: Benjamin
authors.initials: B
authors.lastname: Misselwitz
authors.forename: Sabrina
authors.initials: S
authors.lastname: Dilling
authors.forename: Marcus
authors.initials: M
authors.lastname: Kremer
authors.forename: Wolf-Dietrich
authors.initials: WD
authors.lastname: Hardt
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism*; D019556:COS Cells; D002522:Chlorocebus aethiops; D003092:Colitis / Q000382:microbiology; D003600:Cytosol / Q000382:microbiology; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D008840:Microfilament Proteins / Q000737:chemistry* / Q000378:metabolism*; D008853:Microscopy; D041681:NIH 3T3 Cells; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D012475:Salmonella / Q000166:cytology / Q000472:pathogenicity*
chemical_list: D001426:Bacterial Proteins; D008840:Microfilament Proteins; C118287:SipA protein, Salmonella