title: Cooperation of both, the FKBP_N-like and the DSBA-like, domains is necessary for the correct function of FTS_1067 protein involved in Francisella tularensis virulence and pathogenesis.
abstract: Francisella tularensis the etiological agent of tularaemia is one of the most infectious human pathogen known. Our knowledge about its key virulence factors has increased recently but it still remains a lot to explore. One of the described essential virulence factors is membrane lipoprotein FTS_1067 (nomenclature of F. tularensis subsp. holarctica strain FSC200) with homology to the protein family of disulphide oxidoreductases DsbA. Lipoprotein consists of two different domains: the C-terminal DsbA_Com1-like domain (DSBA-like) and the N-terminal FKBP-type peptidyl-prolyl cis/trans isomerases (FKBP_N-like). To uncover the biological role of these domains, we created bacterial strain with deletion of the DSBA-like domain. This defect in gene coding for lipoprotein FTS_1067 led to high in vivo attenuation associated with the ability to induce host protective immunity. Analyses performed with the truncated recombinant protein showed that the absence of DSBA-like domain revealed the loss of thiol/disulphide oxidoreductase activity and, additionally, confirmed the role of the FKBP_N-like domain in the FTS_1067 oligomerization and chaperone-like function. Finally, we verified that only full-length form of FTS_1067 recombinant protein possesses the isomerase activity. Based on our results, we proposed that for the correct FTS_1067 protein function both domains are needed.
authors.affiliation: Faculty of Military Health Sciences, University of Defence, Department of Molecular Pathology and Biology, Trebesska 1575, Hradec Kralove 50001, Czech Republic.
authors.forename: Iva
authors.initials: I
authors.lastname: Senitkova
authors.affiliation: Faculty of Military Health Sciences, University of Defence, Department of Molecular Pathology and Biology, Trebesska 1575, Hradec Kralove 50001, Czech Republic petra.spidlova@unob.cz.
authors.forename: Petra
authors.initials: P
authors.lastname: Spidlova
authors.affiliation: Faculty of Military Health Sciences, University of Defence, Department of Molecular Pathology and Biology, Trebesska 1575, Hradec Kralove 50001, Czech Republic.
authors.forename: Jiri
authors.initials: J
authors.lastname: Stulik
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D004195:Disease Models, Animal; D005260:Female; D005604:Francisella tularensis / Q000235:genetics / Q000254:growth & development / Q000472:pathogenicity*; D008074:Lipoproteins / Q000235:genetics / Q000378:metabolism; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D008807:Mice, Inbred BALB C; D050505:Mutant Proteins / Q000235:genetics / Q000378:metabolism; D017384:Sequence Deletion; D014406:Tularemia / Q000382:microbiology* / Q000473:pathology; D014774:Virulence; D037521:Virulence Factors / Q000235:genetics / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D008074:Lipoproteins; D008565:Membrane Proteins; D050505:Mutant Proteins; D037521:Virulence Factors
keywords: chaperone; copper ions; oligomerization; protein disulphide isomerase; virulence factor