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Studies on Regulatory Mechanism of Salmonella Pathogenesis by Proteome Analysis

Research Project

Project/Area Number 13470058
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Bacteriology (including Mycology)
Research InstitutionChiba University

Principal Investigator

YAMAMOTO Tomoko  Chiba University, Graduate School of Pharmaceutical Sciences, Professor, 大学院・薬学研究院, 教授 (60110342)

Co-Investigator(Kenkyū-buntansha) TOMOYASU Toshifumi  Chiba University, Graduate School of Pharmaceutical Sciences, Associate Professor, 大学院・薬学研究院, 助教授 (20323404)
TAKAYA Akiko  Chiba University, Graduate School of Pharmaceutical Sciences, Research Assistant, 大学院・薬学研究院, 助手 (80334217)
Project Period (FY) 2001 – 2004
Project Status Completed (Fiscal Year 2004)
Budget Amount *help
¥14,300,000 (Direct Cost: ¥14,300,000)
Fiscal Year 2004: ¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 2003: ¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 2002: ¥4,900,000 (Direct Cost: ¥4,900,000)
Fiscal Year 2001: ¥5,200,000 (Direct Cost: ¥5,200,000)
KeywordsSalmonella / Proteome / Macrophage / Pathogenesis / Apoptosis / Molecular Chaperone / Lon / ClpXP / SPI1 / Lonプロテアーゼ / ClpXPプロテアーゼ / 侵入性 / 鞭毛 / ストレス蛋白質
Research Abstract

Intracellular pathogens, including Salmonella, which maintain long-term resistance within host phagocytes elicit a variety of genetic programs to help them adapt to the hostile environmental conditions encountered within the phagosome. To elucidate the genetic programs for Salmonella pathogenesis expressed in host after infection, a proteome analysis of proteins newly induced in Salmonella after phagocytosis by macrophages was performed. Since prominent among these programs was the heat shock response, the, role of molecular chaperones and AAA+ proteases which are members of the heat shock proteins as virulence proteins have been studied.
(1)The disruption of ClpXP or Lon of serovar Typhimurium results a loss of virulence to mice. The attenuation could be due to the impaired ability to survive and replicate within macrophage cells, suggesting that ClpXP and Lon are critically important for the systemic Salmonella infection of mice. We found that the ClpXP mutant and Lon mutant persist i … More n the BALB/c mice for long periods of time without causing an overwhelming systemic infection, suggesting a possible candidate of Salmonella live vaccine. We therefore examined whether oral immunization with the lon mutant or clpXP mutant protects mice against subsequent oral challenge with virulent serovar Typhimurium. The results suggested that a single oral immunization of the lon and clpXP mutants should be effective to protect against the colonization of wild-type serovar Typhimurium within the intestinal tract.
(2)We demonstrated that depletion of ATP-dependent Lon protease in serovar Typhimurium induces rapid and massive apoptosis in macrophages by a mechanism involving both caspases-1 and -3. This excessive induction of apoptosis was abrogated by disruption of invF, which is required for the expression of the Salmonella pathogenicity island 1(SPI1) genes. Expression of hilA, a central regulator of SPI1 transcription, was repressed in the macrophages after phagocytosis, but this gene was continuously expressed when the Lon mutant grew within the macrophages, so the SPI1 proteins accumulated. Thus, the increase in macrophage apoptosis induced by the Lon mutant could be due to continued expression of SPI1 genes under conditions where they are normally repressed. Once Salmonella has established a systemic infection, excess apoptosis of macrophages cells upon which the organism is reliant would be detrimental to the pathogen. Therefore, the Lon protease may be required to suppress apoptosis sufficiently to allow time for the bacterium to replicate, escape, and invade new macrophages.
(3)The DnaK/DnaJ-depleted mutant lost the ability to causes a lethal systemic disease in mice. Macrophage-survival assays revealed that the mutant could not survive or proliferate at all within macrophages. Of further interest are the findings that the mutant could neither invade cultured epithelial cells nor secrete any of the invasion proteins encoded by Salmonella pathogenicity island 1. Less

Report

(5 results)
  • 2004 Annual Research Report   Final Research Report Summary
  • 2003 Annual Research Report
  • 2002 Annual Research Report
  • 2001 Annual Research Report
  • Research Products

    (45 results)

All 2005 2004 2003 2002 2001 Other

All Journal Article (24 results) Book (6 results) Publications (15 results)

  • [Journal Article] 総説:サルモネラの細胞侵入と食細胞内寄生の分子機構2005

    • Author(s)
      山本友子, 高屋明子
    • Journal Title

      日本細菌学雑誌 60(印刷中)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Degradation of the HilC and HilD regulator proteins by ATP-dependent Lonprotease leads to downregulation of Salmonella pathogenicity island 1 gene expression2005

    • Author(s)
      A.Takaya, Y.Kubota, E.Isogai, T.Yamamoto
    • Journal Title

      Molecular Microbiology 55

      Pages: 839-852

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Derepression of Salmonella pathogenicity island 1 genes within macrophages leads to rapid apoptosis via caspase-1- and caspase-3-dependent pathways2005

    • Author(s)
      A.Takaya, A.Suzuki, Y.Kikuchi, M.Eguchi, E.Isogai, T.Tomoyasu, T.Yamamoto
    • Journal Title

      Cellular Microbiology 7

      Pages: 79-90

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Derepression of Salmonella pathogenicity island 1 genes within macrophages leads to rapid apoptosis via caspase-1-and caspase-3-dependent pathways.2005

    • Author(s)
      A.Takaya, A.Suzuki, Y.Kikuchi, M.Eguchi, E.Isogai, T.Tomoyasu, T.Yamamoto
    • Journal Title

      Cellular Microbiology 7

      Pages: 79-90

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Degradation of the HilC and HilD regulator proteins by ATP-dependent Lon protease leads to down-regulation of Salmonella pathogenicity island 1 gene expression.2005

    • Author(s)
      A.Takaya, Y.Kubota, E.Isogai, T.Yamamoto
    • Journal Title

      Molecular Microbiology 55

      Pages: 839-852

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Degradation of the HilC and HilD regulator proteins by ATP-dependent Lon protease leads to downregulation of Salmonella pathogenicity island 1 gene expression2005

    • Author(s)
      A.Takaya, Y.Kubota, E.Isogai, T.Yamamoto
    • Journal Title

      Molecular Microbiology 55(3)

      Pages: 839-852

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Derepression of Salmonella pathogenicity island 1 genes within macrophages leads to rapid apoptosis via caspase-1- and caspase-3-dependent pathways2005

    • Author(s)
      A.Takaya, A.Suzuki, Y.Kikuchi, M.Eguchi, E.Isogai, T.Tomoyasu, T.Yamamoto
    • Journal Title

      Cellular Microbiology 7(1)

      Pages: 79-90

    • Related Report
      2004 Annual Research Report
  • [Journal Article] The DnaK-DnaJ chaperone machinery of Salmonella enterica serovar Typhimurium is essential for invasion of epithelial cells and survival in macrophages, leading to systemic infection.2004

    • Author(s)
      A.Takaya, T.Tomoyasu, H.Matsui, T.Yamamoto
    • Journal Title

      Infection and Immunity 72

      Pages: 1364-1373

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] The DnaK-DnaJ chaperone machinery of Salmonella enterica serovar Typhimurium is essential for invasion of epithelial cells and survival in macrophages, leading to systemic infection.2004

    • Author(s)
      A.Takaya, T.Tomoyasu, H.Matsui, T.Yamamoto
    • Journal Title

      Infection and Immunity 72(3)

      Pages: 1364-1373

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Lon, a stress induced A ATP-dependent protease is critically important for the systemic Salmonella infection of mice.2003

    • Author(s)
      A.Takaya, M.Suzuki, H.Matsui, T.Tomoyasu, H.Sashinami, A.Nakane, T.Yamamoto
    • Journal Title

      Infection and Immunity 71

      Pages: 690-696

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Oral immunization with ATP-dependent protease deficient mutants protects mice against subsequent oral challenge with virulent Salmonella enterica serovar Typhimurium.2003

    • Author(s)
      H.Matsui, M.Suzuki, Y.Isshiki, C.Kodama, M.Eguchi, Y.Kikuchi, K.Motokawa, A.Takaya, T.Tomoyasu, T.Yamamoto
    • Journal Title

      Infection and Immunity 71

      Pages: 30-39

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] A new heat shock gene agsA, which encodes a small chaperone involved in suppressing protein aggregation in salmonella entrica serovar Typhimurium2003

    • Author(s)
      T.Tomoyasu, A.Takaya, T.Sasaki, T.Nagase, R.Kikuno, M.Morioka, T.Yamamoto
    • Journal Title

      Journal of Bacteriology 185

      Pages: 6331-6339

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Turnover of FlhD and FlhC, masterregulator proteins for Salmonella flagellum biogenesis, by the ATP dependent ClpXP protease2003

    • Author(s)
      T.Tomoyasu, A.Takaya, E.Isogai, T.Yamamoto
    • Journal Title

      Molecular Microbiology 48

      Pages: 443-452

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] A new heat-shock gene-agsA, which encodes a small chaperone involved in suppressing protein aggregation in Salmonella enterica serovar Typhimurium.2003

    • Author(s)
      T.Tomoyasu, A.Takaya, T.Sasaki, T.Nagase, R.Kikuno, M.Morioka T.Yamamoto
    • Journal Title

      Journal of Bacteriology 185

      Pages: 6331-6339

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Turnover of FlhD and FlhC, master regulator proteins for Salmonella flagellum biogenesis, by the ATP-dependent ClpXP protease.2003

    • Author(s)
      T.Tomoyasu, A.Takaya, E.Isogai, T.Yamamoto
    • Journal Title

      Molecular Microbiology 48

      Pages: 443-452

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Lon, a stress-induced ATP-dependent protease, is critically important for the systemic Salmonella enterica serovar Typhamurium infection of mice.2003

    • Author(s)
      A.Takaya, M.Suzuki, H.Matsui, T.Tomoyasu, H.Sashinami, A.Nakane, T.Yamamoto
    • Journal Title

      Infection and Immunity 71

      Pages: 690-696

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Oral immunization with ATP-dependent protease-deficient mutants protects mice against subsequent oral challenge with virulent Salmonella enterica serovar Typhimurium.2003

    • Author(s)
      H.Matsui, M.Suzuki, Y.Isshiki, M.Eguchi, T.Kikuchi, K.Motokawa, A.Takaya, T.Tomoyasu, T.Yamamoto
    • Journal Title

      Infection and Immunity 71

      Pages: 30-39

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] The ClpXP ATP-dependent protease regulates flagellum Synthesis in Salmonella enterica serovar Typhimurium2002

    • Author(s)
      T.Tomoyasu, T.Ohkishi, Y.Ukyo, A.Tokumitsu, A.Takaya, M.Suzuki, K.Sekiya, H.Matsui, K.Kutsukake, T.Yamamoto.
    • Journal Title

      Journal of Bacteriology 184

      Pages: 645-653

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] The ATP-dependent Lon protease of Salmonella enterica serovar Typhimurium regulates invasion and expression of genes carried on Salmonella pathogenicity island 12002

    • Author(s)
      A.Takaya, T.Tomoyasu, A.Tokumitsu, M.Morioka, T.Yamamoto
    • Journal Title

      Journal of Bacteriology 184

      Pages: 224-232

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] 総説:大腸菌のトリガーファクター2002

    • Author(s)
      友安俊文
    • Journal Title

      日本細菌学雑誌 57

      Pages: 597-605

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] The ClpXP ATP-dependent protease regulates flagellum Synthesis in Salmonella enterica serovar Typhimurium.2002

    • Author(s)
      T.Tomoyasu, T.Ohkishi, Y.Ukyo, A.Tokumitsu, A.Takaya, M.Suzuki, K.Sekiya, H.Matsui, K.Kutsukake, T.Yamamoto
    • Journal Title

      Journal of Bacteriology 184

      Pages: 645-653

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] The ATP-dependent Lon protease of Salmonella enterica serovar Typhimurium regulates invasion and expression of genes carried on Salmonella pathogenicity island 1.2002

    • Author(s)
      A.Takaya, T.Tomoyasu, A.Tokumitsu, M.Morioka, T.Yamamoto
    • Journal Title

      Journal of Bacteriology 184

      Pages: 224-232

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Disruption of the genes for ClpXP protease in Salmonella enterica serovar Typhimurium results in persistent infection in mice, and development of persistence requires endogenous gamma interferon and tumour necrosis factor alpha2001

    • Author(s)
      T.Yamamoto, H.Sashinami, A.Takaya, T.Tomoyasu, H.Matsui, T.Hanawa, S.Kamiya, A.Nakane
    • Journal Title

      Infection and Immunity 69

      Pages: 3164-3174

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Disruption of the genes for ClpXP protease in Salmonella enterica serovar Typhimurium results in persistent infection in mice, and development of persistence requires endogenous gamma interferon and tumour necrosis factor alpha.2001

    • Author(s)
      T.Yamamoto, H.Sashinami, A.Takaya, T.Tomoyasu, H.Matsui, T.Hanawa, S.Kamiya, A.Nakane
    • Journal Title

      Infection and Immunity 69

      Pages: 3164-3174

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Book] 感染症学:微生物・寄生虫とのかかわり2005

    • Author(s)
      神谷 茂, 山本友子 他
    • Total Pages
      360
    • Publisher
      日本看護協会出版社
    • Related Report
      2004 Annual Research Report
  • [Book] 薬科微生物学2003

    • Author(s)
      山本 友子(分担執筆)
    • Total Pages
      286
    • Publisher
      丸善株式会社
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Book] 医学大辞典2003

    • Author(s)
      山本 友子(分担執筆)
    • Total Pages
      3062
    • Publisher
      医学書院
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Book] 新世紀の感染症学2003

    • Author(s)
      山本 友子(分担執筆)
    • Total Pages
      837
    • Publisher
      日本臨床
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Book] 微生物学・免疫学2002

    • Author(s)
      山本 友子(分担執筆)
    • Total Pages
      419
    • Publisher
      医学評論社
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Book] 21世紀の考える薬学微生物学2002

    • Author(s)
      山本 友子(分担執筆)
    • Total Pages
      484
    • Publisher
      廣川書店
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Publications] Takaya, A., Tomoyasu, T., Mtasui, H., Yamamoto, T.: "The DnaK-DnaJ chaperone machinery of Salmonella enterica serovar Typhimurium is essential for invasion of epithelial cells and survival in macrophages, leading to systemic infection."Infection and Immunity. 72. 1364-1373 (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] Takaya, A., Suzuki, M., Matsui, H., Tomoyasu, T.Sashinami, H., Nakane, A..Yamamoto, T.: "Lon, a stress induced ATP dependent protease is critically important for the systemic Salmonella infection of mice."Infection and Immunity. 71. 690-696 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Matsui, H., Suzuki, M., Isshiki, Y., Kodama, C., Eguchi, M., Kikuchi, Y., Motokawa, K., Takaya, A., Tomoyasu, T., Yamamoto, T.: "Oral immunization with ATP-dependent protease deficient mutants protects mice against subsequent oral challenge with virulent Salmonella enterica serovar Typhimurium."Infection and Immunity. 71. 30-39 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Tomoyasu, T., Takaya, A., Sasaki, T., Nagase, T., Kikuno, R., Morioka, M., Yamamoto, T.: "A new heat shock gene agsA, which encodes a small chaperone involved in suppressing protein aggregation in Salmonella entrica serovar Typhimurium."Journal of Bacteriology. 185. 6331-6339 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Tomoyasu, T., Takaya, A., Isogai, E., Yamamoto, T.: "Turnover of FlhD and FlhC, masterregulator proteins for Salmonella flagellum biogenesis, by the ATP dependent ClpXP protease."Molecular Microbiology. 48. 443-452 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] 山本 友子: "薬科微生物学"丸善株式会社. 286 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Tomoyasu T. et al.: "Turnover of FlhD and FlhC, master regulator proteins for Salmonella flagellum biogenesis, by the ATP-dependent ClpXP protease"Molecular Microbiology. 48(in press). (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Takaya A., et al.: "Lon, a stress-induced ATP-dependent protease is critically important for the systemic Salmonella infection of mice"Infection and Immunity. 71. 690-696 (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Matsui H., et al.: "Oral immunization with ATP-dependent protease-dificient mutants protects mice against subsequent oral challenge with virulent Salmonella enterica"Infection and Immunity. 71. 31-39 (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Tomoyasu T., et al.: "The ClpXP ATP-dependent protease regulates flagellum synthesis in Salmonella enterica serovar Typhimurium"Journal of Bacteriology. 184. 645-653 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Takaya A., et al.: "The ATP-dependent Lon protease of Salmonella enterica serovar Typhimurium regulates invasion and expression of genes encoded on Salmonella pathogenicity island"Journal of Bacteriology. 184. 224-232 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] 山本友子: "新世紀の感染症学"日本臨床杜. 835 (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Yamamoto, T., H.Sashinami, A.Takaya, T.Tomoyasu, H.Matsui, Y.Kikuchi, T.Hanawa, S.Kamiya, A.Nakane.: "Disruption of the genes for ClpXP protease in Salmonella enterica serovar Typhimurium results in persistence requires endogenous gamma interferon and tumor necrosis factor alpha"Infect.Immun.. 69(5). 3164-3174 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Takaya, A., T.Tomoyasu, A.Tokumitsu, M.Morioka, T.Yamamoto.: "The ATP-dependent Lon protease of Salmonella enterica serovar Typhimurium regulates invasion and expression of genes encoded on Salmonella pathogenicity island 1"J.Bacteriol.. 184(1). 224-232 (2002)

    • Related Report
      2001 Annual Research Report
  • [Publications] Tomoyasu, T., T.Ohkishi, Y.Ukyo, A.Tokumitsu, A.Takaya, M.Suzuki, K.Sekiya, H.Matsui, K.Kutsukake, T.Yamamoto: "The ClpXP ATP-dependent protease regulates flagellum synthesis in Salmonella enterica serovar Typhimurium"J.Bacteriol.. 184(3). 645-653 (2002)

    • Related Report
      2001 Annual Research Report

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Published: 2001-04-01   Modified: 2016-04-21  

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