Functional analysis of XAB2 complex involved in transcription and DNA repair

2002 Fiscal Year Final Research Report Summary

• Project/Area Number: 13680763
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Molecular biology
• Research Institution: Kyushu University
• Principal Investigator: NAKATSU Yoshimichi Kyushu University, Granduate Schools, Faculty of Medical Science, Associate Professor, 医学研究院, 助教授 (00207820)
• Project Period (FY): 2001 – 2002
• Keywords: nucreotide excision repair / transcription / TCR / XPA / XAB2

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.

Research Products

• [Publications] Yoshino M, Nakatsu Y, Riele H, Hirota S, Kitamura Y, Tanaka K.: "Additive roles of XPA and MSH2 genes in UVB-induced skin tumorigenesis in mice"DNA Repair. 1・11. 935-940 (2002)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Murai M, Enokido Y, Inamura N, Yoshino M, Nakatsu Y, van der Horst GT, Hoeijmakers JH, Tanaka K, Hatanaka H.: "Early postnatal ataxia and abnormal cerebellar development in mice lacking Xeroderma pigmentosum Group A and Cockayne syndrome Group B DNA repair genes"Proc.Natl.Acad.Sci.USA. 98・23. 13379-13384 (2001)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Tanaka K, Kamiuchi S, Ren Y, Yonemasu R, Ichikawa M, Murai H, Yoshino M, Takeuchi S, Saijo M, Nakatau Y, Miyauchi-Hashimoto H, Horio T.: "UV-induced skin carcinogenesis in xeroderma pigmentosum group A (XPA) gene-knockout mice with nucleotide excision repair-deficiency"Mutation Research. 477・1-2. 31-40 (2001)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Yoshino M., Nakatsu Y., Riele H., Hirota S., Kitamura Y., Tanaka K.: "Additive roles of XPA and MSH2 genes in UVB-induced skin tumorigenesis in mice"DNA Repair. 1-11. 935-940 (2002)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Murai M., Enokido Y., Inamura N., Yoshino M, Nakatsu Y., van der Horst GT., Hoeijmakers JH., Tanaka K., Hatanaka H.: "Early postnatal ataxja and abnormal cerebellar development in mice lacking Xeroderma pigmentosum Group A and Cockayne syndrome Group B DNA repair genes"Proc. Natl. Acad. Sci. U S A. 98-23. 13379-13384 (2001)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Tanaka K., Kamiuchi S., Ren Y., Yonemasu R., Ichikawa,M., Murai,H., Yoshino,M., Takeuchi,S., Saijo,M., Nakatsu,Y., Miyauchi-Hashimoto H., Horio T.: "UV-induced skin carcinogenesis in xeroderma pigmentosum group A (XPA) gene-knockout mice with nucleotide excision repair-deficiency"Mutation Research. 477-1-2. 31-40 (2001)
  • Description: 「研究成果報告書概要(欧文)」より