title: Pivotal role of the Francisella tularensis heat-shock sigma factor RpoH.
abstract: Francisella tularensis is a highly infectious pathogen that infects animals and humans to cause the disease tularemia. The primary targets of this bacterium are macrophages, in which it replicates in the cytoplasm after escaping the initial phagosomal compartment. The ability to replicate within macrophages relies on the tightly regulated expression of a series of genes. One of the most commonly used means of coordinating the regulation of multiple genes in bacteria consists of the association of dedicated alternative sigma factors with the core of the RNA polymerase (RNAP). In silico analysis of the F. tularensis LVS genome led us to identify, in addition to the genes encoding the RNAP core (comprising the alpha1, alpha2, beta, beta' and omega subunits), one gene (designated rpoD) encoding the major sigma factor sigma(70), and a unique gene (FTL_0851) encoding a putative alternative sigma factor homologue of the sigma(32) heat-shock family (designated rpoH). Hence, F. tularensis represents one of the minority of bacterial species that possess only one or no alternative sigma factor in addition to the main factor sigma(70). In the present work, we show that FTL_0851 encodes a genuine sigma(32) factor. Transcriptomic analyses of the F. tularensis LVS heat-stress response allowed the identification of a series of orthologues of known heat-shock genes (including those for Hsp40, GroEL, GroES, DnaK, DnaJ, GrpE, ClpB and ClpP) and a number of genes implicated in Francisella virulence. A bioinformatic analysis was used to identify genes preceded by a putative sigma(32)-binding site, revealing both similarities to and differences from RpoH-mediated gene expression in Escherichia coli. Our results suggest that RpoH is an essential protein of F. tularensis, and positively regulates a subset of genes involved in the heat-shock response.
authors.affiliation: INSERM, U570, Unit of Pathogenesis of Systemic Infections, F-75015 Paris, France.|Université Paris Descartes, Faculté de Médecine Necker-Enfants Malades, F-75015 Paris, France.
authors.forename: Nathalie
authors.initials: N
authors.lastname: Grall
authors.affiliation: Channing Laboratories, Brigham and Women's Hospital, Harvard Medical School, 181 Longwood Avenue, Boston, MA 02115, USA.
authors.forename: Jonathan
authors.initials: J
authors.lastname: Livny
authors.affiliation: Howard Hughes Medical Institute, 4000 Jones Bridge Road, Chevy Chase, MD 20815-6789, USA.|Channing Laboratories, Brigham and Women's Hospital, Harvard Medical School, 181 Longwood Avenue, Boston, MA 02115, USA.
authors.forename: Matthew
authors.initials: M
authors.lastname: Waldor
authors.affiliation: INSERM, U570, Unit of Pathogenesis of Systemic Infections, F-75015 Paris, France.|Université Paris Descartes, Faculté de Médecine Necker-Enfants Malades, F-75015 Paris, France.
authors.forename: Monique
authors.initials: M
authors.lastname: Barel
authors.affiliation: INSERM, U570, Unit of Pathogenesis of Systemic Infections, F-75015 Paris, France.|Université Paris Descartes, Faculté de Médecine Necker-Enfants Malades, F-75015 Paris, France.
authors.forename: Alain
authors.initials: A
authors.lastname: Charbit
authors.affiliation: INSERM, U570, Unit of Pathogenesis of Systemic Infections, F-75015 Paris, France.|Université Paris Descartes, Faculté de Médecine Necker-Enfants Malades, F-75015 Paris, France.
authors.forename: Karin L
authors.initials: KL
authors.lastname: Meibom
mesh_terms: D016384:Consensus Sequence; D004269:DNA, Bacterial / Q000032:analysis / Q000235:genetics; D004926:Escherichia coli / Q000378:metabolism; D005604:Francisella tularensis / Q000235:genetics / Q000378:metabolism* / Q000472:pathogenicity*; D020869:Gene Expression Profiling; D015964:Gene Expression Regulation, Bacterial; D005816:Genetic Complementation Test; D016680:Genome, Bacterial; D006360:Heat-Shock Proteins / Q000737:chemistry / Q000502:physiology*; D018869:Heat-Shock Response; D012808:Sigma Factor / Q000737:chemistry / Q000502:physiology*; D014158:Transcription, Genetic; D014774:Virulence
chemical_list: D004269:DNA, Bacterial; D006360:Heat-Shock Proteins; D012808:Sigma Factor; C069774:heat-shock sigma factor 32