| Project/Area Number |
11640642
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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 |
植物生理
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| Research Institution | Tokyo University of Agriculture and Technology |
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Principal Investigator |
FUKUGARA Toshiyuki Tokyo.University of Agriculture and Technology, Faculty of Agriculture, Associate professor, 農学部, 助教授 (90228924)
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| Project Period (FY) |
1999 – 2001
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2001: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 2000: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1999: ¥1,200,000 (Direct Cost: ¥1,200,000)
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| Keywords | Double-stranded RNA (dsRNA) / DsRNA-binding protein / DsRNA-binding domain / Arabidopsis thaliana / Westwestern / Northwestern / Rice / Protein-proteininteraction / 形質転換体 / 遺伝子破壞株 / ノースウエタン法 |
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| Research Abstract |
We have found 12 putative genes for double-stranded RNA (dsKNA)-binding proteins (DRBPs) in Arabidopsis thaliana, all of which contain the conserved dsRNA-binding domain (DRBD) consisting of about 70 amino acid residues. We have characterized five putative DRBPs : ADB1, ADB2 and ADB3 from A. thaliana and ODB1 from rice that are about 400 amino acid residues in length with two DRBDs at their N-terminal regions, and CAP that has two putative RNaselH domains in the N-terminal region and two putative DRBDs in the C-terminal regions.
By Northwestern assays, the recombinant proteins of these five genes had the binding activity specific for dsRNAs, but strength of their binding activities were different to each other. The dsRNA-binding activity ofADB2 was weaker than those of the other four proteins. Furthermore, their binding activites to 100 bp dsRNAwas similarto those to 500 bp dsRNAbutthose to 25 bp dsRNA was very weak.
Interaction between these DRBPs were analyzed by Westwestern assay. ADB 1 interacted with CAF via its DRBDs and their binding activity between ADB1 and CAP was greater than its between ADB1 and ADB1. ADB2 and ADB3 also bound to ADB1, CAF and themselves. These DRBPs may interact to each other in vivo.
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