1998 Fiscal Year Final Research Report Summary
ANALYSIS OF TRANSCRIPTIONAL REGULATION OF RECOMBINATION ACTIVATING GENE (RAG) DURING PROCESS OF LYMOHOID DEVEROPMENT
| Project/Area Number |
09836004
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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 |
免疫の制御機構
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| Research Institution | TOYAMA MEDICAL AND PHARMACEUTICAL UNIVERSITY |
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Principal Investigator |
MURAGUCHI Atsushi TOYAMA MED AND PHAR.UNIV., FACULTY OF MEDICEN,Professor, 医学部, 教授 (20174287)
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| Co-Investigator(Kenkyū-buntansha) |
KISHI Hiroyuki TOYAMA MED AND PHAR.UNIV., FACULTY OF MEDICEN,Associate Professor, 医学部, 助教授 (60186210)
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| Project Period (FY) |
1997 – 1998
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| Keywords | T CELL RECEPTOR / IMMUNOGLOBULIN / GENE REARRANGEMENT / RECOMBINASE / RAG / PROMOTOR / TRANSCRIPTION FACTOR / CIS-ELEMNT |
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| Research Abstract |
1. Reseach for human rag genomic locus Using a fragment of human rag-1 cDNA, we isolated genomic DNA clones that contained either first or second exon of human rag-1 . We determined its exon/intron organization and the transcriptional start site by the primer extenstion analysis and RNA protection assay. Similarly. we isolated genomic DNA clones that contains first or second exon of human rag-2, using a probe obtained from a fragment of human rag-2 cDNA, and determined its exon/intron structure and the transcription initiation site for rag-2. These led us to reveal, for the first time, the organization of human genomic rag-1 and rag-2 locus.
2. Research for regulation of rag transcription ; We characterized promoter regions for both rag- 1 and rag-2. By transient expression assays using a luciferase reporter gene with truncated promoter fragments, or substitution mutants, we showed that the 5 promoter region containing the ccaat-box between -110 and -86 was indispensable for its basal promoter activity. On contrary, in case of rag-2 promoter, the sequences between -63 to -107, which contained neither ccaat-box or reported cis-element, were shown to be essential for its promoter acticity, suggesting the regulation of rag-1 and rag-2 transcription may be controled by different transcriptional mechanisms. Now, we are trying to determine the cis-element and isolate the transcription factor that binds to the cis- element and regulates the transcription of rag-2.
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