| Project/Area Number |
16570141
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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 |
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Principal Investigator |
NAGAWA Fumikiyo The University of Tokyo, Graduate School of Science, Lecturer, 大学院・理学系研究科, 講師 (10241233)
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| Co-Investigator(Kenkyū-buntansha) |
NISHIZUMI Hirofumi The University of Tokyo, Graduate School of Science, Research Associate, 大学院・理学系研究科, 助手 (30292832)
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| Project Period (FY) |
2004 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| Budget Amount *help |
¥3,800,000 (Direct Cost: ¥3,800,000)
Fiscal Year 2005: ¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 2004: ¥2,000,000 (Direct Cost: ¥2,000,000)
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| Keywords | antigen receptors / V(D)J recombination / RAG proteins / V(D)J組換え / フットプリント解析 / 3'末端プロセシング |
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| Research Abstract |
In V(D)J joining of antigen receptor genes, two recombination signal sequences, 12- and 23-RSSs, form a synaptic complex with the protein products of recombination activating genes, RAG1 and RAG2. In order to study the interaction between the RAG proteins and the RSS DNAs during the process of V(D)J joining, we purified the pre- and post-cleavage type complexes in vitro and analyzed them by DNase I footprinting. We found that the interaction of RAG proteins with substrate RSS DNA is not just limited to the signal region but it extends into the coding region at least up to the 12th nucleotide from the RSS-coding border. Joining mutants of RAG1 and RAG2 demonstrate impaired interactions with the coding region in both pre- and post-cleavage type complexes. Although the exact nature of the mutants' defect is still unknown, the impaired interaction with the coding sequence in the post-cleavage complex may be reason for the joining defect observed with mutant proteins. Our results suggest that the coding-region interaction with the RAG proteins is important not only in the pre-cleavage phase, but also in the post-cleavage process of V(D)J recombination.
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