2016 Fiscal Year Annual Research Report
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
|
|---|
| Research Institution | Nagoya University |
|---|
Principal Investigator |
荻 朋男 名古屋大学, 環境医学研究所, 教授 (80508317)
|
|---|
| 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
|
|---|
| Project Period (FY) |
2014-04-25 – 2017-03-31
|
| Keywords | TC-NER / RNA polymerase Ⅱ / Ubiquitination / UV sensitive syndrome |
|---|
| 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.
|
| Research Progress Status |
28年度が最終年度であるため、記入しない。
|
| Strategy for Future Research Activity |
28年度が最終年度であるため、記入しない。
|
-
[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.
-
Journal Title
Journal of Dermatology
Volume: 印刷中
Pages: 印刷中
DOI
Peer Reviewed / Open Access
-
-
-
[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.
-
Journal Title
Photodermatol Photoimmunol Photomed
Volume: 32
Pages: 174-180
DOI
Peer Reviewed / Open Access / Int'l Joint Research
-
[Presentation] DNA修復機構の異常と疾患.2017
Author(s)
荻朋男
Organizer
太陽紫外線防御研究委員会第27回シンポジウム
Place of Presentation
佐治敬三メモリアルホール（大阪府大阪市）
Year and Date
2017-03-17 – 2017-03-17
Invited
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
