title: Francisella tularensis subsp. holarctica DsbA homologue: a thioredoxin-like protein with chaperone function.
abstract: Francisella tularensis is a highly infectious facultative intracellular bacterium and aetiological agent of tularaemia. The conserved hypothetical lipoprotein with homology to thiol/disulphide oxidoreductase proteins (FtDsbA) is an essential virulence factor in F. tularensis. Its protein sequence has two different domains: the DsbA_Com1_like domain (DSBA), with the highly conserved catalytically active site CXXC and cis-proline residue; and the domain amino-terminal to FKBP-type peptidyl-prolyl isomerases (FKBP_N). To establish the role of both domains in tularaemia infection models, site-directed and deletion mutagenesis affecting the active site (AXXA), the cis-proline (P286T) and the FKBP_N domain (ΔFKBP_N) were performed. The generated mutations led to high attenuation with the ability to induce full or partial host protective immunity. Recombinant protein analysis revealed that the active site CXXC as well as the cis-proline residue and the FKBP_N domain are necessary for correct thiol/disulphide oxidoreductase activity. By contrast, only the DSBA domain (and not the FKBP_N domain) seems to be responsible for the in vitro chaperone activity of the FtDsbA protein.
authors.affiliation: Institute of Molecular Pathology, Faculty of Military Health Sciences, University of Defence, 500 01 Hradec Kralove, Czech Republic.|Department of Biochemical Studies, Faculty of Pharmacy in Hradec Kralove, Charles University in Prague, 500 05 Hradec Kralove, Czech Republic.
authors.forename: Monika
authors.initials: M
authors.lastname: Schmidt
authors.affiliation: Institute of Molecular Pathology, Faculty of Military Health Sciences, University of Defence, 500 01 Hradec Kralove, Czech Republic.
authors.forename: Jana
authors.initials: J
authors.lastname: Klimentova
authors.affiliation: Institute of Molecular Pathology, Faculty of Military Health Sciences, University of Defence, 500 01 Hradec Kralove, Czech Republic.
authors.forename: Pavel
authors.initials: P
authors.lastname: Rehulka
authors.affiliation: Center of Advanced Studies, Faculty of Military Health Sciences, University of Defence, 500 01 Hradec Kralove, Czech Republic.
authors.forename: Adela
authors.initials: A
authors.lastname: Straskova
authors.affiliation: Institute of Molecular Pathology, Faculty of Military Health Sciences, University of Defence, 500 01 Hradec Kralove, Czech Republic.
authors.forename: Petra
authors.initials: P
authors.lastname: Spidlova
authors.affiliation: Department of Biochemical Studies, Faculty of Pharmacy in Hradec Kralove, Charles University in Prague, 500 05 Hradec Kralove, Czech Republic.
authors.forename: Barbora
authors.initials: B
authors.lastname: Szotakova
authors.affiliation: Institute of Molecular Pathology, Faculty of Military Health Sciences, University of Defence, 500 01 Hradec Kralove, Czech Republic.
authors.forename: Jiri
authors.initials: J
authors.lastname: Stulik
authors.affiliation: Institute of Molecular Pathology, Faculty of Military Health Sciences, University of Defence, 500 01 Hradec Kralove, Czech Republic.
authors.forename: Ivona
authors.initials: I
authors.lastname: Pavkova
mesh_terms: D004252:DNA Mutational Analysis; D005604:Francisella tularensis / Q000201:enzymology* / Q000235:genetics*; D018832:Molecular Chaperones / Q000235:genetics* / Q000378:metabolism*; D016297:Mutagenesis, Site-Directed; D050505:Mutant Proteins / Q000235:genetics / Q000378:metabolism; D019704:Protein Disulfide-Isomerases / Q000235:genetics* / Q000378:metabolism*; D017384:Sequence Deletion; D013879:Thioredoxins / Q000235:genetics / Q000378:metabolism; D014774:Virulence; D037521:Virulence Factors / Q000235:genetics / Q000378:metabolism
chemical_list: D018832:Molecular Chaperones; D050505:Mutant Proteins; D037521:Virulence Factors; D013879:Thioredoxins; D019704:Protein Disulfide-Isomerases