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Molecular analysis of anteliotic-tolerance and stress response in adherent bacteria

Research Project

Project/Area Number 17591914
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Morphological basic dentistry
Research InstitutionThe University of Tokushima

Principal Investigator

ONO Tsuneko  The University of Tokushima, School of Medicine, Professor (40035514)

Co-Investigator(Kenkyū-buntansha) MIYAKE Yoichiro  The University of Tokushima, Institute of Health Biosciences, Graduate School, Professor (80136093)
ABIKO Yoshimitsu  Nihon University, School of Dentistry at Matsudo, Professor (70050086)
HIGUCHI Tomihiko  The University of Tokushima, Institute of Health Biosciences, Graduate School, professor (50035557)
KUWAHARA Tomomi  The University of Tokushima, Institute of Health Biosciences, Graduate School, Associate Professor (60263810)
KAYAMA Shizuo  The University of Tokushima, Institute of Health Biosciences, Graduate School, Assistant Professor (50432761)
村上 圭史  徳島大学, 大学院ヘルスバイオサイエンス研究部, 助手 (10335804)
Project Period (FY) 2005 – 2007
Project Status Completed (Fiscal Year 2007)
Budget Amount *help
¥3,610,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥210,000)
Fiscal Year 2007: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2006: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 2005: ¥2,000,000 (Direct Cost: ¥2,000,000)
Keywordsantibiotic tolerance / stress response / rpoS / adherent bacteria / ppGppc / rpoN / pvdS / vqsR / quorum sensing / 遺伝子発現 / pyoverdine / pvdS / vqsR / tolerance / 抗菌薬 / tcp gene / Pseudomonas aeruginosa
Research Abstract

In this project, we have studied about the molecular mechanism of antibiotic tolerance and stress response in adherent cells The following results werte obtained in this study.
1. To assess the contribution of ppGpp in antibiotic tolerance to quinolone in Pseudomonas aeruginosa, knockout mutants of relA, spoT and dksA, were constructed and investigated for their antibiotic susceptibility to quinolones. The survival of the dksA and spoT mutants in the presence of ofloxacin and ciprofloxacin were shown to be approximately 20-180 and 10-40 times respectively, higher than the same for the wild type strain.
The data suggest that elevated basal levels of ppGpp may be responsible for rendering these mutants tolerant to quinolones and expand the importance of ppGpp as an antimicrobial target in P. aeruginosa
2. An rpoN mutant in Pseudomonas aeruginosa has been constructed and investigated its importance as a target for antibiotics. The stationary phase cells of the rpoN mutant displayed an approximately 15 times higher survival rate to quinolones and carbapenems than the wild-type cells. Real-time PCR revealed that in contrast to the wild-type strain, the stationary phase cells of the rpoN mutant grown without antibiotics had approximately 4-140 and 2-14-fold higher transcripts of pvdS and vqsR genes, respectively, than the wild type strain. In the presence of antibiotics, pvdS and vqsR transcripts were 400 and 5-fold, respectively, elevated in comparison to the wild-type. Flow cytometry assays using GFP reporter demonstrated increased expression of the vqsR gene in the rpoN mutant. A pvdS mutant of P. aeruginosa, deficient in pyoverdine production, was shown to be susceptible to biapenem.
These findings suggest that rpoN is involved in the tolerance to antimicrobial agents in P. aeruginosa and its tolerant effect is partly dependent on increased pyoverdine production and vqsR gene expression.

Report

(4 results)
  • 2007 Annual Research Report   Final Research Report Summary
  • 2006 Annual Research Report
  • 2005 Annual Research Report
  • Research Products

    (42 results)

All 2008 2007 2006 2005 Other

All Journal Article (11 results) (of which Peer Reviewed: 2 results) Presentation (31 results)

  • [Journal Article] rpoN gene of Pseudomonas aeruginosa alters its susceptibility to quinolones and carbapenems2007

    • Author(s)
      D. Viducic
    • Journal Title

      Antimicrobiol. Agents Chemother 51

      Pages: 1455-1462

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] 付着状態における緑膿菌の抗菌薬抵抗性とQuorum sensing機構について2007

    • Author(s)
      鹿山鎭男
    • Journal Title

      Bacterial Adherence & Biofilm 21(印刷中)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] rpoN gene of Pseudomonas aeruginosa alters its susceptibility to quinolones and carbapenems2007

    • Author(s)
      D. Viducic, et. al.
    • Journal Title

      Antimicrobiol. Agents Chemother 51

      Pages: 1455-1462

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Role for Quorum Sensing Related Genes of Adherent Pseudomonas aeruginosa in Antibiotic Tolerance2007

    • Author(s)
      S. Kayama, et. al.
    • Journal Title

      Bacterial Adherence & Biofilm 21(in press)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] 付着状態における緑膿菌の抗菌薬抵抗性とQuorum sensing機構について2007

    • Author(s)
      鹿山 鎮男
    • Journal Title

      Bacterial Adherence & Biofilm 21

      Pages: 49-53

    • Related Report
      2007 Annual Research Report
  • [Journal Article] rpoN Gene of Pseudomonas aeruginosa Alters Its Susceptibility to Quinolones and Carbapenems2007

    • Author(s)
      Darija Viducic
    • Journal Title

      Antimicrobiol.Agents Chemother. 51・4

      Pages: 1455-1462

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Function Analysis of spoT, relA and dksA genes on quinolone tolerance in Pseudomonas aeruginosa under nongrowing condition.2006

    • Author(s)
      D. Viducic
    • Journal Title

      Microbiol. Immunol 50

      Pages: 349-357

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] 付着緑膿菌の抗菌薬抵抗性とQuorum sensing機構について2006

    • Author(s)
      S. Kayama鹿山鎭男
    • Journal Title

      Bacterial Adherence & Biofilm 20

      Pages: 110-115

    • NAID

      10019310355

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Function Analysis of spoT, relA and dksA genes on quinolone tolerance in Pseudomonas aeruginosa under nongrowing condition2006

    • Author(s)
      D. Viducic, et. al.
    • Journal Title

      Microbiol. Immunol 50

      Pages: 349-357

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Role for Quorum Sensing Related Genes of Adherent Pseudomonas aeruginosa in Antibiotic Tolerance2006

    • Author(s)
      S. Kayama, et. al.
    • Journal Title

      Bacterial Adherence & Biofilm 20

      Pages: 110-115

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Functional analysis of spoT, relA and dksA genes on quinolone tolerance in quinolone tolerance in Pseudomonas aeruginosa under nongrowing condition2006

    • Author(s)
      Darija Viducic
    • Journal Title

      Microbiol. Immmunol 50・4(印刷中)

    • Related Report
      2005 Annual Research Report
  • [Presentation] A role of ppGpp in tolerance to quinolone in Pseudomonas aeruginosa2008

    • Author(s)
      D. Viducic, et. al.
    • Organizer
      Pseudomonas 10th International Congress
    • Place of Presentation
      Marseille
    • Year and Date
      2008-08-28
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Alteration of intracellular Ca^<2+> concentration and non-apoptotic cell death induced by intermedilysin Intermedilysin2008

    • Author(s)
      K. Hirota
    • Organizer
      The 81th Annual Meeting of Japanese Society for Bacteriology
    • Place of Presentation
      Kyoto
    • Year and Date
      2008-05-26
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Gene Expression and Antibiotic tolerance of adherent cells in Pseudomonas aeruginosa2008

    • Author(s)
      S. Kayama
    • Organizer
      The 81th Annual Meeting of Japanese Society for Bacteriology
    • Place of Presentation
      Kyoto
    • Year and Date
      2008-05-25
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Intermedilysinにより誘導される非アポトーシス型細胞死と細胞内Ca^<2+>濃度の変動2008

    • Author(s)
      弘田 克彦
    • Organizer
      第81回日本細菌学会総会
    • Place of Presentation
      京都
    • Year and Date
      2008-03-26
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] 付着状態における緑膿菌の抗菌薬抵抗性と遺伝子発現について2008

    • Author(s)
      S. Kayama鹿山 鎭男
    • Organizer
      第81回日本細菌学会総会
    • Place of Presentation
      京都
    • Year and Date
      2008-03-25
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Annual Research Report 2007 Final Research Report Summary
  • [Presentation] 付着緑膿菌の抗菌薬抵抗性とQuorum Sensing機構について2007

    • Author(s)
      鹿山 鎭男
    • Organizer
      第55回日本化学療法学会西日本支部総会
    • Place of Presentation
      神戸
    • Year and Date
      2007-10-31
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Quorum sensing system and antimicrobial tolerance in adherent cells in Pseudomonas aeruginosa2007

    • Author(s)
      S. Kayama
    • Organizer
      The 55th Annual Meeting of Western Japan Branch in Japanese Society for Chemotherapy
    • Place of Presentation
      Kobe
    • Year and Date
      2007-10-31
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] 付着状態における緑膿菌の抗菌薬抵抗性とquorum sensing機構について2007

    • Author(s)
      鹿山 鎮男
    • Organizer
      第55回日本化学療法学会西日本支部総会
    • Place of Presentation
      神戸
    • Year and Date
      2007-10-31
    • Related Report
      2007 Annual Research Report
  • [Presentation] Role of quorum sensing system of adherent Pseudomonas aeruginosa in antibiotic tolerance2007

    • Author(s)
      Shizuo, Kayama
    • Organizer
      ASM conferences Pseudomonas
    • Place of Presentation
      Seattle
    • Year and Date
      2007-08-28
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Role of quorum sensing system of adherent Pseudomonas aeruginosa in antibiotic tolerance2007

    • Author(s)
      鹿山 鎮男
    • Organizer
      ASM Conferences Pseudomonas 2007
    • Place of Presentation
      Seattle,USA
    • Year and Date
      2007-08-28
    • Related Report
      2007 Annual Research Report
  • [Presentation] 付着状態における緑膿菌の抗菌薬抵抗性とQuorum sensing機構について2007

    • Author(s)
      鹿山 鎭男
    • Organizer
      Bacterial Adherence&Biofilm第21回学術集会
    • Place of Presentation
      東京
    • Year and Date
      2007-07-07
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] 付着緑膿菌の抗菌薬抵抗性とQuorum Sensing機構について2007

    • Author(s)
      鹿山 鎭男
    • Organizer
      第80回日本細菌学会総会
    • Place of Presentation
      大阪
    • Year and Date
      2007-03-28
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Role of quorum sensing system of adherent Pseudomonas aeruginosa in antibiotic tolerance2007

    • Author(s)
      Shizuo Kayama
    • Organizer
      ASM conferences Pseudomonas 2007
    • Place of Presentation
      Seattle
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Role of quorum sensing system to antibiotic tolerance in adherent Pseudomonas aeruginosa2007

    • Author(s)
      S. Kayama
    • Organizer
      The 21th Bacterial Adherence & Biofilm
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] 抗菌薬添加による緑膿菌Quorum Sensing関連遺伝子発現の変化2006

    • Author(s)
      村上 圭史
    • Organizer
      第54回日本化学療法学会西日本支部総会
    • Place of Presentation
      福岡
    • Year and Date
      2006-12-03
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Alteration of rpoS gene expression by antibiotics in Pseudomonas aeruginosa2006

    • Author(s)
      K. Murakami
    • Organizer
      The 54th Annual Meeting of Western Japan Branch in Japanese Society for Chemotherapy
    • Place of Presentation
      Fukuoka
    • Year and Date
      2006-12-03
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Effect of antibiotics on rpoS promoter in Pseudomonas aeruginosa2006

    • Author(s)
      K. Murakami
    • Organizer
      The 79th Annual Meeting of Japanese Society for Bacteriology
    • Place of Presentation
      Kanazawa
    • Year and Date
      2006-05-29
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Antibiotic Tolerance and Quorum Sensing System in Adherent Bacteria2006

    • Author(s)
      S. Kayama
    • Organizer
      The 79th Annual Meeting of Japanese Society for Bacteriology
    • Place of Presentation
      Kanazawa
    • Year and Date
      2006-05-29
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] 抗菌薬添加による緑膿菌rpoS遺伝子発現の変化2006

    • Author(s)
      村上 圭史
    • Organizer
      第54回日本化学療法学会
    • Place of Presentation
      京都
    • Year and Date
      2006-05-19
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Alteration of rpoS gene expression by antibiotics in Pseudomonas aeruginosa2006

    • Author(s)
      K. Murakami
    • Organizer
      The 54th Annual Meeting of Japanese Society for Chemotherapy
    • Place of Presentation
      Kyoto
    • Year and Date
      2006-05-19
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] 付着細菌の抗菌薬抵抗性とQuorum sensing機構について2006

    • Author(s)
      鹿山 鎭男
    • Organizer
      第79回日本細菌学会総会
    • Place of Presentation
      金沢
    • Year and Date
      2006-03-31
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] 抗菌薬による緑膿菌rpoS遺伝子プロモーター活性への影響2006

    • Author(s)
      村上 圭史
    • Organizer
      第79回日本細菌学会総会
    • Place of Presentation
      金沢
    • Year and Date
      2006-03-29
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] 付着緑膿菌の抗菌薬抵抗性とQuorum sensing機構について2006

    • Author(s)
      鹿山 鎭男
    • Organizer
      Bacterial Adherence&Biofilm第20回記念学術集会
    • Place of Presentation
      東京
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Antibiotic Tolerance and Quorum Sensing System in adherent Bacteria2006

    • Author(s)
      S. Kayama
    • Organizer
      The 20th Bacterial Adherence & Biofilm
    • Place of Presentation
      Tokyo
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] 緑膿菌Quorum Sensingとofloxacin抵抗性について2005

    • Author(s)
      鹿山 鎭男
    • Organizer
      第53回日本化学療法学会西日本支部総会
    • Place of Presentation
      岐阜
    • Year and Date
      2005-12-02
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Quorum sensing and ofloxacin tolerance in Psudomonas aeruginosa2005

    • Author(s)
      S. Kayama
    • Organizer
      The 53th Annual Meeting of Western Japan Branch in Japanese Society for Chemotherapy
    • Place of Presentation
      Gifu
    • Year and Date
      2005-12-02
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] The involvement of pslL gene in biapenem tolerance of Pseudomonas aeruginosa2005

    • Author(s)
      K Murakami
    • Organizer
      Pseudomonas 10th International Congress
    • Place of Presentation
      Marseille
    • Year and Date
      2005-08-30
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] A role of ppGpp in tolerance to quinolone in Pseudomonas aeruginosa2005

    • Author(s)
      D. Viducic
    • Organizer
      Pseudomonas 10th International Congress
    • Place of Presentation
      Marseille
    • Year and Date
      2005-08-28
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] 緑膿菌tcp遺伝子が抗菌薬抵抗性及ぼす影響について2005

    • Author(s)
      村上 圭史
    • Organizer
      第58回日本細菌学会中国四国支部総会
    • Place of Presentation
      高知
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Effect of tcp gene of Pseudomonas aeruginosa on antimicrobial tolerance2005

    • Author(s)
      K. Murakami
    • Organizer
      The 58th Annual Meeting of Japanese Society for Bacteriology Chugoku-Shikoku Branch
    • Place of Presentation
      Kochi
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Quorum sensing System and Antibiotic Tolerance in adherent Pseudomonas aeruginosa

    • Author(s)
      S. Kayama
    • Organizer
      The 80th Annual Meeting of Japanese Society for Bacteriology
    • Place of Presentation
      Osaka
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary

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Published: 2005-04-01   Modified: 2016-04-21  

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