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

2010 Fiscal Year Final Research Report

• Project/Area Number: 21780088
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Single-year Grants
• Research Field: Applied biochemistry
• Research Institution: Tohoku 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
• Keywords: 細胞応答 / 自然免疫 / 抗菌ペプチド

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.

Research Products

• [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
  • 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
  • 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
• [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
• [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
• [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
• [Remarks] ホームページ等