title: TheHelicobacter pyloriJ99jhp0106Gene, under the Control of the CsrA/RpoN Regulatory System, Modulates Flagella Formation and Motility.
abstract: CsrA has been shown to positively control the expression of flagella-related genes, includingflaAandflaB, through regulating expression of an alternative sigma factor RpoN inHelicobacter pyloriJ99. Here, we aimed to characterize the CsrA regulatory system by comparative transcriptomic analysis carried out with RNA-seq on strain J99 and acsrAmutant. Fifty-three genes in thecsrAmutant were found to be differentially expressed compared with the wild-type. Among CsrA-regulated genes,jhp0106, with unclear function, was found located downstream offlaBin the J99 genome. We hypothesized thatflaB-jhp0106is in an operon under the control of RpoN binding to theflaBpromoter. The RT-qPCR results showed the expression ofjhp0106was decreased 76 and 92% in thecsrAandrpoNmutants, respectively, compared to the wild-type. Moreover, mutations of the RpoN binding site in theflaBpromoter region resulted in decreased expression offlaBandjhp0106and deficient motility. Three-dimensional structure modeling results suggested that Jhp0106 was a glycosyltransferase. The role ofjhp0106inH. pyloriwas further investigated by constructing thejhp0106mutant and revertant strains. A soft-agar motility assay and transmission electron microscope were used to determine the motility and flagellar structure of examined strains, and the results showed the loss of motility and flagellar structure injhp0106mutant J99. In conclusion, we foundjhp0106, under the control of the CsrA/RpoN regulatory system, plays a critical role inH. pyloriflagella formation.
authors.affiliation: Department of Biotechnology and Laboratory Science in Medicine, School of Biomedical Science and Engineering, National Yang Ming University Taipei, Taiwan.
authors.forename: Cheng-Yen
authors.initials: CY
authors.lastname: Kao
authors.affiliation: Center of Infectious Disease and Signaling Research, National Cheng Kung UniversityTainan, Taiwan; Department of Microbiology and Immunology, College of Medicine, National Cheng Kung UniversityTainan, Taiwan.
authors.forename: Jenn-Wei
authors.initials: JW
authors.lastname: Chen
authors.affiliation: Center of Infectious Disease and Signaling Research, National Cheng Kung UniversityTainan, Taiwan; Department of Microbiology and Immunology, College of Medicine, National Cheng Kung UniversityTainan, Taiwan.
authors.forename: Shuying
authors.initials: S
authors.lastname: Wang
authors.affiliation: Department of Internal Medicine, College of Medicine, National Cheng Kung University Hospital, National Cheng Kung UniversityTainan, Taiwan; Department of Internal Medicine, Tainan Hospital, Ministry of Health & WelfareTainan, Taiwan.
authors.forename: Bor-Shyang
authors.initials: BS
authors.lastname: Sheu
authors.affiliation: Department of Biotechnology and Laboratory Science in Medicine, School of Biomedical Science and Engineering, National Yang Ming University Taipei, Taiwan.
authors.forename: Jiunn-Jong
authors.initials: JJ
authors.lastname: Wu
keywords: CsrA; flagella; glycosylation; jhp0106; motility