転写共役修復開始反応の分子機能解析（遺伝性光線過敏症の分子基盤）
| Project/Area Number |
14F04093
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| Research Category |
Grant-in-Aid for JSPS Fellows
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| Allocation Type | Single-year Grants |
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| Section | 外国 |
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| Research Field |
Risk sciences of radiation and chemicals
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| Research Institution | Nagoya University (2015-2016)
Nagasaki University (2014) |
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Principal Investigator |
荻 朋男 名古屋大学, 環境医学研究所, 教授 (80508317)
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| Co-Investigator(Kenkyū-buntansha) |
GUO CHAOWAN 名古屋大学, 環境医学研究所, 外国人特別研究員
GUO Chaowan 名古屋大学, 環境医学研究所, 外国人特別研究員
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| Project Period (FY) |
2014-04-25 – 2017-03-31
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| Project Status |
Completed (Fiscal Year 2016)
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| Budget Amount *help |
¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 2016: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 2015: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 2014: ¥700,000 (Direct Cost: ¥700,000)
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| Keywords | TC-NER / RNA polymerase Ⅱ / Ubiquitination / UV sensitive syndrome / RNA polymerase II / Cockyane syndrome / ヌクレオチド除去修復機構 / UVSSA / 紫外線高感受性症候群 / RNAポリメラーゼII / ユビキチン化 |
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| Outline of Annual Research Achievements |
We firstly generated a cell line library, in which each TC-NER factors (e.g. UVSSA or Usp7) is knocked out using CRISPR/Cas9 technique; To get insights into UVSSA-dependent RNA polIIo ubiquitination, we setup a SILAC-based quantitative proteomics method for efficiently isolation and enrichment of ubiquitinated peptides, in combination with high-resolution mass-spectrum and we successfully identified numbers of UV-damage specific and UVSSA-dependent ubiquitination sites on stalled RNA polIIo; To subsequently test the biological relevance of polIIo ubiquitination to TC-NER, the “non-ubiquitinable” mutant cell lines were generated and several generated mutant cell lines displayed reduced TC-NER activity and defects in RNA polIIo ubiquitination upon UV irradiation. To give a detail view of various TC-NER proteins in RNA polIIo ubiquitination, we have purified most of TC-NER factors including RNA polII, UVSSA, CSA/B, Cul4CSA E3 ligase and Usp7. By using these purified proteins, we set out to reproduce the RNA polIIo ubiquitination, a smear of RNA polIIo modification were observed by addition of RNA polII and Cul4CSA E3 ligase in reaction, which was previously reported. But after several attempts, so far we could not reproduce the UVSSA-specific RNA polII ubiquitination in test tube, implying that this reaction requires further factor (s) to complete. We are now setting up an proteomic method to identify potential factors that may involved in this process, and we believe the following work will provide further information for molecular basis of RNA polIIo ubiquitination.
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| Research Progress Status |
28年度が最終年度であるため、記入しない。
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| Strategy for Future Research Activity |
28年度が最終年度であるため、記入しない。
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Report
(3 results)
Research Products
(32 results)
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[Journal Article] Analysis of clinical symptoms and ABCC6 mutations in 76 Japanese patients with pseudoxanthoma elasticum.2017
Author(s)
Iwanaga A, Okubo Y, Yozaki M, Koike Y, Kuwatsuka Y, Tomimura S, Yamamoto Y, Tamura H, Ikeda S, Maemura K, Tsuiki E, Kitaoka T, Endo Y, Mishima H, Yoshiura KI, Ogi T, Tanizaki H, Wataya-Kaneda M, Hattori T, Utani A.
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Journal Title
Journal of Dermatology
Volume: 印刷中
Issue: 6
Pages: 644-650
DOI
Related Report
Peer Reviewed / Open Access / Int'l Joint Research
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[Journal Article] A ten-year follow up of a child with mild case of xeroderma pigmentosum complementation group D diagnosed by whole genome sequencing.2016
Author(s)
Ono R, Masaki T, Mayca Pozo F, Nakazawa Y, Swagemakers SM, Nakano E, Sakai W, Takeuchi S, Kanda F, Ogi T, van der Spek PJ, Sugasawa K, Nishigori C.
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Journal Title
Photodermatol Photoimmunol Photomed.
Volume: 未定
Issue: 4
Pages: 174-180
DOI
Related Report
Peer Reviewed / Open Access / Int'l Joint Research
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[Journal Article] XRCC4 deficiency in human subjects causes a marked neurological phenotype but no overt immunodeficiency.2015
Author(s)
Guo C, Nakazawa Y, Woodbine L, Bjorkman A, Shimada M, Fawcett H, Jia N, Ohyama K, Li TS,Nagayama Y, Mitsutake N, Pan-Hammarström Q, Gennery AR, Lehmann AR, Jeggo PA, Ogi T.
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Journal Title
J Allergy Clin Immunol.
Volume: 136 (4)
Issue: 4
Pages: 1007-1017
DOI
Related Report
Peer Reviewed / Open Access / Int'l Joint Research
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[Presentation] DNA損傷応答と遺伝病疾患2016
Author(s)
荻朋男
Organizer
平成28年度若手放射線生物学研究会専門研究会
Place of Presentation
東京工業大学（東京都目黒区）
Year and Date
2016-09-03
Related Report
Invited
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