| Project/Area Number |
13680763
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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 | Kyushu University |
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Principal Investigator |
NAKATSU Yoshimichi Kyushu University, Granduate Schools, Faculty of Medical Science, Associate Professor, 医学研究院, 助教授 (00207820)
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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 |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2002: ¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 2001: ¥1,900,000 (Direct Cost: ¥1,900,000)
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| Keywords | nucreotide excision repair / transcription / TCR / XPA / XAB2 / RNAポリメラーゼII / 遺伝子欠損マウス / CSA / CSB / 転写 / PPIE |
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| Research Abstract |
XPA-binding protein 2 (XAB2) is involved in transcription and transcription-coupled repair (TCR), and interacts with the TCR factors, CSA, CSB and RNA polymerase II. Flag-tagged XAB2 was expressed in HeLa cells and purified with ant-Flag antibody. Flag-tagged XAB2 was co-purified with about 20 polypeptides. Consistent with previous observation that microinjected anti-XAB2 antibody inhibits transcription and TCR, this affinity purified XAB2 complex enhanced FACT activity that stimulates an in vitro transcription elongation reaction using a chromatin template. Further purification of this protein complex with gel-filtration and miniQ column chromatography indicated that XAB2 was tightly associated with 5 polypeptides: p160, p57, p50, p42, and p35. Analysis using the mass spectrography revealed that p35 is peptidy1 proly1 cis-trans isomerase E (PPIE), a rotamase with RNA binding activity. Yeast 2-hybrid assay and in vitro pull-down assay with several deletion mutant proteins of PPIE indicated that XAB2 is directly associated with PPIE and RNA binding domain of PPIE is essential for the interaction.
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