Molecular mechanism and biological meaning of transmitting in pre-mRNA of the transcription factor Sp1
| Project/Area Number |
13680684
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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 |
Structural biochemistry
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| Research Institution | The University of Tokyo |
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Principal Investigator |
AKANUMA Hiroshi The University of Tokyo, Graduate School of Aits and Sciences , Professor, 大学院・総合文化研究科, 教授 (30012462)
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| Co-Investigator(Kenkyū-buntansha) |
YANAGISAWA Shuichi The University ofTokyo, Graduate School ofAife and Sciences, Assistant Professor, 大学院・総合文化研究科, 助手 (20222359)
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| Project Period (FY) |
2001 – 2002
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| Project Status |
Completed (Fiscal Year 2002)
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| Budget Amount *help |
¥3,700,000 (Direct Cost: ¥3,700,000)
Fiscal Year 2002: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 2001: ¥2,800,000 (Direct Cost: ¥2,800,000)
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| Keywords | tans-splicing / splicing / transcription factor / Sp1 / トランス・スプライシング / スプライシング / トランス・スプライシンズ |
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| Research Abstract |
Recently, we showed that trans-splicing between pre-mRNAs from the gene for the human transcription factor, Sp1 produces the variants of the mRNAin cultured cells. Although several examples for trans-splicing between pre-mRNA derived from the same gene have recently reported, the molecular mechanism for this type of trans-splicing remains largely unknown. We proceeded to this project to reveal the molecular mechanism for trans-splicing between pre-mRNA derived from a single gene and to search the biological meaning of trans-splicing between Sp1 pre-mKNAs. We hypothesized that the presence of a large intron in Sp1 gene (intron 3) might cause the trans-splicing. Because splicing is coupled with transcription, the donor site of a large intron might be spliced with the acceptor site within the following pre-mRNA, before synthesis of the acceptor site for cis-splicing. To verify this possibility, we transformed human HepG2 cells and obtained several stable transformants harboring the Sp1 genes with modified intron-exon structures. The PCR-based analyzes for trans-splicing in transformants revealed that the length rather than the DNA-sequence of the intron 3 was a critical factor for trans-splicing between Sp1 pre-mRNAs. In the other hand, we found that the trans-splicing between Sp1 pre-mRNAs occurred in rats in the tissue-independent manner. The ratio of the trans-spliced products to the cis-spliced product was approximately 1%. Splicing machinery in mammalian cells might involve the activity for trans-spli'cing as a side reaction and such activity might be graetely enhanced by the presence of large introns that increase free donor sites.
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Report
(3 results)
Research Products
(3 results)
