Identification of novel defense factors using Luciferase expressing Listeria by RNAi screening.
| Project/Area Number |
21780088
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| Research Category |
Grant-in-Aid for Young Scientists (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Applied biochemistry
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| Research Institution | Tohoku University |
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Principal Investigator |
GOTO Akira Tohoku University, 大学院・生命科学研究科, 助教 (70419000)
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| Project Period (FY) |
2009 – 2010
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| Project Status |
Completed (Fiscal Year 2010)
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| 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)
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| 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 |
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| 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.
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Report
(3 results)
Research Products
(16 results)
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[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
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[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
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