2001 Fiscal Year Final Research Report Summary
The analysis of general transcription machinary by RNA aptamer
| Project/Area Number |
12680673
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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 | KYOTO UNIVERSITY |
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Principal Investigator |
HIRAYOSHI Kazunori Inst. for Frontier Medical Sciences, Kyoto Univ., Lecturer, 再生医科学研究所, 講師 (80199108)
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| Project Period (FY) |
2000 – 2001
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| Keywords | RNA aptamer / pre-initiation complex / surface plasmon resonance / SELEX / TBP |
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| Research Abstract |
We introduced a new molecule RNA aptamer to analyze the mechanism of the establishment of pre-initiation complex, paused polymerase as a model. RNA aptamer would work in the cell, (i) like an antibody, be target a particular protein, (ii) like a small organic molecule, be able to deliver within cells, and (iii) like a conditional allele, be able to exert its effect in whole organisms. We selected an RNA aptamer against to TBP (TATA binding protein), which is a key molecule for establishment of pre-initiation complex. The dissociation constant of selected RNA aptamers are between 10^<-7> to 10^<-8> by Surface plasmon resonance analysis. These numbers show specific binding of RNA aptamer to TBP. Selected RNA aptamers are divided to two groups. One can stimulate the transcription and the other can inhibit the transcription and accumulate the short transcript. These results show that the combination of general transcription factors centered on TBP regulate the transcription. The binding sites of each RNA aptamer will clarify the architecture of transcription complex. After the analyses of binding property of RNA aptamer, we will move into in vivo analysis of the transcription using RNA aptamers.
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