Biochemical analysis of the AGO-siRNA complex about DNA binding activity to induce the TGS (Transcriptional Gene Silencing).
| Project/Area Number |
21570178
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Molecular biology
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| Research Institution | The University of Tokyo (2010-2011)
Waseda University (2009) |
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Principal Investigator |
ISHIDA Takaomi 東京大学, 医科学研究所, 特任准教授 (80293447)
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| Project Period (FY) |
2009 – 2011
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| Project Status |
Completed (Fiscal Year 2011)
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| Budget Amount *help |
¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000)
Fiscal Year 2011: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2010: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2009: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
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| Keywords | 転写 / TGS / Gene Silencing / Argonaute / epigenetics / siRNA / アルゴノート / 転写調節 / gene silencing / HIV |
|---|
| Research Abstract |
Our research group previously published data demonstrate that of siRNA targeting conserved regions of the untranscribed HIV-1 promoter region, a single dose of either Prom A siRNA induced prolonged and profound suppression of HIV re-production. Definitive evidence of transcriptional silencing (TGS) was provided to this repression of HIV reproduction by several assays. There is accumulating evidence the Argonaute family of siRNA binding proteins, play a pivotal role in siRNA induced TGS. In TGS Ago1 protein accumulates at the site of initiation of silencing. To explore the role of Ago1 in siRNA induced TGS of HIV-1, in vitro binding assay was established for detecting AGO1-siRNA complex can be bind to target DNA. This assay system clearly demonstrated that these complex canb ebind directly to taget DNA sequence. AGO1 protein was co-localized with siRNA when the cells infected HIV-1 by immune-staining assay. We also demonstrated that these TGS effects are dependent upon close matching between the siRNA and the target sequence.
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Report
(4 results)
Research Products
(28 results)
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[Journal Article] Viral interference with host mRNA surveillance, the nonsense-mediated mRNA decay (NMD) pathway, through a new function of HTLV-1 Rex: implicationsfor retroviral replication.2013
Author(s)
Nakano K, Ando T, Yamagishi M, Yokoyama K, Ishida T, Ohsugi T, Tanaka Y, Brighty DW, Watanabe T.
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Journal Title
Related Report
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[Journal Article] Transcriptional gene silencing of HIV-1 through promoter targeted RNA is highly specific.2011
Author(s)
Suzuki K, Ishida T, Yamagishi M, Ahlenstiel C, Swaminathan S, Marks K, Murray D, McCartney EM, Beard MR, Alexander M, Purcell DF, Cooper DA, Watanabe T, Kelleher AD.
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Journal Title
RNA Biol.
Volume: 8(6)
Pages: 1035-46
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[Presentation] Promoter targeting small RNA suppresses HIV-1 infection in vivo.2013
Author(s)
Kazuo Suzuki, S Hattori, K Marks, C Ahlenstiel, M Millington, M Boyd, G Symonds, Takaomi Ishida, D Cooper, Seiji Okada, A Kelleher.
Organizer
20^<th> Conference on retroviruses and opportunistic infections 2013.
Place of Presentation
Atlanta, USA.
Related Report
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[Presentation] Promoter targeting small RNA suppresses HIV-1 infection in vivo.2013
Author(s)
Kazuo Suzuki, S Hattori, K Marks, C Ahlenstiel, M Millington, M Boyd, G Symonds, Takaomi Ishida, D Cooper, Seiji Okada, A Kelleher.
Organizer
20th Conference on retroviruses and opportunistic infections 2013.
Place of Presentation
Atlanta, USA
Related Report
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[Presentation] The Epgenetic Regulation of HIV-1 virus Gene Expression.2010
Author(s)
Takaomi Ishida, Kazuo Suzuki, Makoto Yamagishi, David A Cooper, Anthony D. Lelleher, Kentaro Semba, Tadashi Yamamoto,Aikichi Iwamoto and Toshiki Watanabe.
Organizer
The 7th Japan-China joint laboratory workshop pathogenesis, gene regulation, and signal transduction.
Place of Presentation
Beijing, China.
Year and Date
2010-10-25
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