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Identification of novel defense factors using Luciferase expressing Listeria by RNAi screening.

Research Project

Project/Area Number 21780088
Research Category

Grant-in-Aid for Young Scientists (B)

Allocation TypeSingle-year Grants
Research Field Applied biochemistry
Research InstitutionTohoku University

Principal Investigator

GOTO Akira  Tohoku University, 大学院・生命科学研究科, 助教 (70419000)

Project Period (FY) 2009 – 2010
Project Status Completed (Fiscal Year 2010)
Budget Amount *help
¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000)
Fiscal Year 2010: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
Fiscal Year 2009: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
Keywords細胞応答 / 自然免疫 / 抗菌ペプチド / DNAマイクロアレイ / RNAi / ショウジョウバエ / 細胞内寄生細菌 / レポーターアッセイ / コロニー形成法 / リステリア菌 / DNAマイクロアレイ法
Research Abstract

Innate immune system is one of the first-line defense system that is highly conserved from plants, insects and mammals. Peptidoglycan recognition protein (PGRP)-LE firstly identified in our laboratory is functioning on both intracellular and extracellular space and responsible for the activation of one of the innate immune signaling pathway, Imd pathway, and melanin synthesis pathway. Furthermore, PGRP-LE plays a crucial role to eliminate intracellular pathogens using one of the general protein degradation system called "Autophagy", which is independent on the Toll and Imd pathway. Therefore, to identify further novel host defense factors invoved in this multi-functional PGRP-LE, I set up the system where the growth of Listeria can be monitored by constitutive Luciferase expressing Listeria strains. Using this Listeria strain, the infection condition where we can see the effect of PGRP-LE was scrutinized. As an alternative strategy, I also conducted DNA microarray and identified a novel antimicrobial-like peptide named Listericin. Further analysis of Listericin using protein purification, RNAi and overexpression studies demonstrated that the expression of Listericin is cooperatively regulated by both PGRP-LE and the JAK-STAT pathway, as a secreted form, it exibits bacteriocidal activities against gram-negative bacteria and Listeria strain. Furthermore, in vivo approach using Listericin overexpressing transgenic flies showed that they exhibit resistance against Listeria infection in the surival test. Taken together with these results, we can show that Listericin is a novel antimicrobial peptide on Drosophila innate immune response.

Report

(3 results)
  • 2010 Annual Research Report   Final Research Report ( PDF )
  • 2009 Annual Research Report

Research Products

(16 results)

All 2010 2009 Other

All Journal Article Presentation Remarks

  • [Journal Article] Cooperative regulation of the induction of the novel antibacterial Listericin by PGRP-LE and the JAK-STAT pathway.2010

    • Author(s)
      Akira Goto, Tamaki Yano, Jun Terashima, Shinzo Iwashita, Yoshiteru Oshima, Shoichiro Kurata.
    • Journal Title

      Journal of Biological Chemistry. Vol.285

      Pages: 15731-15738

    • Related Report
      2010 Final Research Report
    • Peer Reviewed
  • [Journal Article] Immune response of insects and crustaceans.2010

    • Author(s)
      Akira Goto, Shoichiro Kurata.
    • Journal Title

      Bentham Science eBook (Review article) Accepted

    • Related Report
      2010 Final Research Report
    • Peer Reviewed
  • [Journal Article] Cooperative regulation of the induction of the novel antibacterial Listericin by PGRP-LE and the JAK-STAT pathway.2010

    • Author(s)
      Akira Goto, Tamaki Yano, Jun Terashima, Shinzo Iwashita, Yoshiteru Oshima, Shoichiro Kurata
    • Journal Title

      Journal of Biological Chemistry

      Volume: VOL.285 Pages: 15731-15738

    • Related Report
      2010 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Immune response of insects and crustaceans.2010

    • Author(s)
      Akira Goto, Shoichiro Kurata
    • Journal Title

      Bentham Science eBook (Review article)

      Volume: (Accepted)

    • Related Report
      2010 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Cooperative regulation of the induction of the novel antibacterial Listericin by PGRP-LE and the JAK-STAT pathway2010

    • Author(s)
      Akira Goto, Tamaki Yano, Jun Terashima, Shinzo Iwashita, Yoshiteru Oshima, Shoichiro Kurata
    • Journal Title

      Journal of Biological Chemistry (Accepted, 印刷中)

    • Related Report
      2009 Annual Research Report
    • Peer Reviewed
  • [Presentation] Novel roles of receptor-type Guanylyl Cyclase Gyc76C on Drosophila innate immune responses.2010

    • Author(s)
      Goto, A., Iwashita, S., Nabe, K., Fukuzaki, M., Oshima, Y., Kurata, S.
    • Organizer
      BMB2010(第33回分子生物学会年会、第83回日本生化学会大会合同大会)(口答発表&ポスター発表)
    • Place of Presentation
      神戸
    • Year and Date
      2010-12-09
    • Related Report
      2010 Final Research Report
  • [Presentation] Cooperative regulation of the induction of the novel antibacterial Listericin by PGRP-LE and JAK-STAT pathway.2010

    • Author(s)
      Goto, A., Kurata, S.
    • Organizer
      14th International Congress of Immunology (第14回国際免疫学会議)
    • Place of Presentation
      神戸
    • Year and Date
      2010-08-23
    • Related Report
      2010 Final Research Report
  • [Presentation] Cooperative regulation of the induction of the novel antibacterial Listericin by PGRP-LE and JAK-STAT pathway.2010

    • Author(s)
      Goto, A., Yano, T., Terashima, J., Iwashita, S., Oshima, Y., Kurata S.
    • Organizer
      日本比較免疫学会第22回学術集会(古田奨励賞受賞)
    • Place of Presentation
      福岡
    • Year and Date
      2010-08-02
    • Related Report
      2010 Final Research Report
  • [Presentation] 新規抗菌ペプチドDrosophila Listericinの同定2010

    • Author(s)
      後藤彰(G-COE助教)、矢野環(准教授)、寺島潤(助教)、倉田祥一朗(教授)
    • Organizer
      第76回日本生化学会東北支部会例会
    • Place of Presentation
      福島
    • Year and Date
      2010-05-08
    • Related Report
      2010 Final Research Report
  • [Presentation] 新規抗菌ペプチドDrosophila Listericinの同定2010

    • Author(s)
      後藤彰(G-COE助教)、矢野環(准教授)、寺島潤(助教)、倉田祥一朗(教授)
    • Organizer
      第76回日本生化学会東北支部会例会
    • Place of Presentation
      福島コラッセ(福島県)
    • Year and Date
      2010-05-08
    • Related Report
      2010 Annual Research Report
  • [Presentation] Cooperative regulation of the induction of the novel antibacterial Listericin by PGRP-LE and JAK-STAY pathway.2010

    • Author(s)
      Akira Goto, Tamaki Yano, Jun Terashima, Shinzo Iwashita, Yoshiteru Oshima, Shoichiro Kurata.
    • Organizer
      日本比較免疫学会第22回学術集会
    • Place of Presentation
      九州大学西新プラザ(福岡県)
    • Related Report
      2010 Annual Research Report
  • [Presentation] Cooperative regulation of the induction of the novel antibacterial Listericin by PGRP-LE and JAK-STAY pathway2010

    • Author(s)
      Akira Goto, Shoichiro Kurata
    • Organizer
      14th International Congress of Immunology(第14回国際免疫学会議)
    • Place of Presentation
      神戸ホートピアホテル、神戸国際展示場(兵庫県)
    • Related Report
      2010 Annual Research Report
  • [Presentation] Novel roles of receptor-type Guanylyl Cyclase Gyc76C on Drosophila innate immune responses.2010

    • Author(s)
      Akira Goto, Shinzo Iwashita, Kumiko Nabe, Masataka Fukuzaki, Yoshiteru Oshima, Shoichiro Kurata.
    • Organizer
      BMB2010(第33回分子生物学会年会、第83回日本生化学会大会 合同大会)
    • Place of Presentation
      神戸ホートピアホテル、神戸国際展示場(兵庫県)
    • Related Report
      2010 Annual Research Report
  • [Presentation] Novel antimicrobial peptide-like gene Listericin induced by Listeria monocytogenes infection has bacteriocidal activities against gram-negative bacteria2009

    • Author(s)
      Akira Goto, Tamaki Yano, Jun Terashima, Shoichiro Kurata
    • Organizer
      JDRC9(The 9th Japanese Drosophila Research Conference)
    • Place of Presentation
      Yamaha Resort TSUMAGO I, Shizuoka, Japan
    • Related Report
      2009 Annual Research Report
  • [Presentation] Novel Drosophila antimicrobial peptide-like gene Listericin regulated by JAK-STAT pathway has bacteriocidal activities against gram-negative bacteria2009

    • Author(s)
      Akira Goto, Tamaki Yano, Jun Terashima, Shoichiro Kurata
    • Organizer
      Woodstoich2009(Ecological/Biological Stoichiometry Workshop for young scientists)
    • Place of Presentation
      Kawatabi Seminar Center of Tohoku University, Kawatabi, Japan
    • Related Report
      2009 Annual Research Report
  • [Remarks] ホームページ等

    • Related Report
      2010 Final Research Report

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Published: 2009-03-31   Modified: 2016-04-21  

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