A rapid, comprehensive system for assaying DNA repair activity and cytotoxic effects of DNA-damaging reagents

• Project/Area Number: 24659533
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Dermatology
• Research Institution: Nagasaki University
• Principal Investigator: OGI Tomoo 長崎大学, 原爆後障害医療研究所, 准教授 (80508317)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000); Fiscal Year 2013: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2012: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
• Keywords: ヌクレオチド除去修復 / DNA修復

Outline of Final Research Achievements

Nucleotide excision repair (NER) is most versatile DNA repair process: it prevents cells from genomic instability as it removes various DNA damages arose in the genome. DNA repair assays that can directly measure NER activities are useful for clinical diagnoses and clinical researches including anticancer drug discovery. Unscheduled DNA synthesis (UDS) and recovery of RNA synthesis (RRS) are two commonly used DNA repair endpoints for assessing the activity of NER. We developed a comprehensive procedure for measuring UDS, RRS and cell-sensitivity by the incorporation of ethynyl-nucleoside analogs (EdU and EU) followed by the click-chemistry reaction.
The developed system is suitable for a virus-based UDS / RRS / cell-sensitivity complementation tests, by which we can systematically determine the pathogenic gene of a patient with defect in the DNA repair pathways. A typical clinical diagnosis, using human primary fibroblast cells, takes less than 1 week.

Research Products

• [Journal Article] A rapid, comprehensive system for assaying DNA repair activity and cytotoxic effects of DNA-damaging reagents. (2015)
  • Author(s): Nan Jia, Yuka Nakazawa, Chaowan Guo, Mayuko Shimada, Mieran Sethi, Yoshito Takahashi, Hiroshi Ueda, Yuji Nagayama, Tomoo Ogi
  • Journal Title: Nature Protocols Volume: 10 Issue: 1 Pages: 12-24
  • DOI: 10.1038/nprot.2014.194
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Hypomorphic PCNA mutation underlies a human DNA repair disorder. (2014)
  • Author(s): Baple EL, Chambers H, Cross HE, Fawcett H, Nakazawa Y, Chioza BA, Harlalka GV, Mansour S, Sreekantan-Nair A, Patton MA, Muggenthaler M, Rich P, Wagner K, Coblentz R, Stein CK, Last JI, Taylor AM, Jackson AP, Ogi T, Lehmann AR, Green CM, Crosby AH.
  • Journal Title: Journal of Clinical Investigation Volume: 124 Issue: 7 Pages: 3137-3146
  • DOI: 10.1172/jci74593
  • Peer Reviewed
• [Journal Article] PRKDC mutations in a SCID patient with profound neurological abnormalities. (2013)
  • Author(s): Woodbine L, Neal JA, Sasi NK, Shimada M, Deem K, Coleman H, Dobyns WB, Ogi T, §Meek K, §Davies EG, §Jeggo PA.
  • Journal Title: The Journal of Clinical Investigation Volume: 123 Issue: 7 Pages: 2969-2980
  • DOI: 10.1172/jci67349
  • Peer Reviewed
• [Journal Article] Malfunction of Nuclease ERCC1-XPF Results in Diverse Clinical Manifestations and Causes Cockayne Syndrome, Xeroderma Pigmentosum, and Fanconi Anemia. (2013)
  • Author(s): Kashiyama, K, Nakazawa, Y, Pilz, DT, Guo, C, Sasaki, K, et al.
  • Journal Title: The American Journal of Human Genetics Volume: 92 Issue: 5 Pages: 807-819
  • DOI: 10.1016/j.ajhg.2013.04.007
  • Peer Reviewed
• [Journal Article] Functional characterization of the novel BRAF complex mutation, BRAF(V600delinsYM), identified in papillary thyroid carcinoma. (2013)
  • Author(s): Matsuse, M.
  • Journal Title: Int. J. Cancer Volume: 132 Issue: 3 Pages: 738-743
  • DOI: 10.1002/ijc.27709
  • Peer Reviewed
• [Journal Article] Identification of the first ATRIP-deficient patient and novel mutations in ATR define a clinical spectrum for ATR-ATRIP Seckel syndrome (2012)
  • Author(s): *Ogi T , Walker S, Stiff T, Hobson E, Limsirichaikul S, Carpenter G, Prescott K, Suri M, Byrd P, Matsuse M, Mitsutake N, Nakazawa Y, Vasudevan P, Barrow M, Stewart G, Taylor M, O'Driscoll M, and Jeggo P
  • Journal Title: PLoS Genetics Volume: 8 Issue: 11 Pages: e1002945-e1002945
  • DOI: 10.1371/journal.pgen.1002945
  • NAID: 120006985789
  • Peer Reviewed
• [Journal Article] Mutations in UVSSA cause UV-sensitive syndrome and impair RNA polymerase IIo processing in transcription-coupled nucleotide-excision repair (2012)
  • Author(s): Nakazawa Y, Sasaki K, Mitsutake N, Yamashita S, Ogi T, et al
  • Journal Title: Nat Genet Volume: 44(5) Issue: 5 Pages: 586-92
  • DOI: 10.1038/ng.2229
  • NAID: 120006985586
  • Peer Reviewed
• [Presentation] 放射線感受性および各種発達異常を示す遺伝性疾患の新規責任遺伝子の同定と分子機能解析 (2014)
  • Author(s): 中沢由華、郭朝万、嶋田繭子、宮崎仁美、唐田清伸、荻朋男
  • Organizer: 第37回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜（神奈川県横浜市）
  • Year and Date: 2014-11-25 – 2014-11-27
• [Presentation] Molecular and functional study on the initiation of transcription coupled nucleo tide excision repair (2014)
  • Author(s): Guo C, Nakazawa Y, Shimada M, Jia N, Karata K, Miyazaki H, Ogi T.
  • Organizer: 第37回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜（神奈川県横浜市）
  • Year and Date: 2014-11-25 – 2014-11-27
• [Presentation] 転写と共役したヌクレオチド除去修復のin vitro反応系の構築 (2014)
  • Author(s): 唐田清伸、郭朝万、荻朋男
  • Organizer: 第37回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜（神奈川県横浜市）
  • Year and Date: 2014-11-25 – 2014-11-27
• [Presentation] コケイン症候群様の臨床症状を示す遺伝性疾患の責任遺伝子探索 (2014)
  • Author(s): 宮崎仁美、中沢由華、郭朝万、嶋田繭子、賈楠、唐田清伸、荻朋男
  • Organizer: 第37回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜（神奈川県横浜市）
  • Year and Date: 2014-11-25 – 2014-11-27
• [Presentation] エキソーム解析を用いたDNA修復機構欠損性疾患の新規責任遺伝子の探索 (2014)
  • Author(s): 嶋田繭子、中沢由華、郭朝万、賈楠、宮崎仁美、唐田清伸、荻朋男
  • Organizer: 第37回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜（神奈川県横浜市）
  • Year and Date: 2014-11-25 – 2014-11-27
• [Presentation] Molecular cloning and characterisation of new human DNA repair genes (2014)
  • Author(s): Ogi T
  • Organizer: The 9th 3R Symposium
  • Place of Presentation: 御殿場高原ホテル（静岡県御殿場市）
  • Year and Date: 2014-11-17 – 2014-11-21
• [Presentation] Molecular characterization and functional analysis of XRCC4, a novel pathological gene for radiation sensitivity and developmental abnormalities (2014)
  • Author(s): Guo C, Nakazawa Y, Shimada M, Woodbine L, Jia N, Karata K, Miyazaki H, Lehmann A, Jeggo PA, Ogi T.
  • Organizer: The 9th 3R Symposium
  • Place of Presentation: 御殿場高原ホテル（静岡県御殿場市）
  • Year and Date: 2014-11-17 – 2014-11-21
• [Presentation] ERCC1/XPF deficiency causes three NER- deficient disorders: a patient with various symptoms of xeroderma pigmentosum, Cockayne syndrome & Fanconi anemia. (2014)
  • Author(s): Nakazawa Y, Kashiyama K, Pilz DT, Guo C, Shimada M, Sasaki K, Fawcett H, Wing JF, Lewin SO, Carr L, Li TS, Yoshiura K, Utani A, Hirano A, Greenblatt D, Nardo T, Stefanini M, McGibbon D, Sarkany R, Fassihi H, Mitsutake N, Lehmann AR, Ogi T.
  • Organizer: The 9th 3R Symposium
  • Place of Presentation: 御殿場高原ホテル（静岡県御殿場市）
  • Year and Date: 2014-11-17 – 2014-11-21
• [Presentation] ゲノム不安定性疾患群の新規責任遺伝子の同定と分子機能解析 (2014)
  • Author(s): 荻朋男
  • Organizer: 第57回日本甲状腺学会学術集会
  • Place of Presentation: コングレコンベンションセンター（大阪府大阪市）
  • Year and Date: 2014-11-14
  • Invited
• [Presentation] ヒストンH3K9メチル化酵素類のDNA二重鎖切断修復反応への関与 (2014)
  • Author(s): 荻朋男
  • Organizer: 第87回日本生化学会大会 シンポジウム
  • Place of Presentation: 国立京都国際会館（京都府京都市）
  • Year and Date: 2014-10-15 – 2014-10-18
• [Presentation] 転写共役ヌクレオチド除去修復の開始反応の分子機構 (2014)
  • Author(s): 荻朋男
  • Organizer: 日本放射線影響学会第57回大会 ワークショップ
  • Place of Presentation: 鹿児島県民交流センター（鹿児島県鹿児島市）
  • Year and Date: 2014-10-01 – 2014-10-03
• [Presentation] DNA修復機構の異常により発症する先天性疾患とゲノム不安定性/発がん (2014)
  • Author(s): 荻朋男
  • Organizer: 第20回日本家族性腫瘍学会学術集会 教育講演
  • Place of Presentation: コラッセ福島（福島県福島市）
  • Year and Date: 2014-06-13
  • Invited
• [Presentation] Transcription, DNA damage and Repair (2014)
  • Author(s): 荻朋男
  • Organizer: International Symposium on Xeroderma Pigmentosum and Related Diseases
  • Place of Presentation: 神戸国際会議場（神戸市）
• [Presentation] 紫外線高感受性症候群～UVSSA遺伝子の発見と病態解析 (2013)
  • Author(s): 荻朋男
  • Organizer: 第22回DNA複製・組換え・修復ワークショップ
  • Place of Presentation: ホテルニュー水戸屋（仙台市）
• [Presentation] Transcription, DNA damage and Repair (2013)
  • Author(s): 荻朋男
  • Organizer: 11th International Conference on Environmental Mutagens
  • Place of Presentation: Bourbon Convention & Spa Resort Cataratas do Iguassu（ブラジル）
  • Invited
• [Presentation] 転写共役修復とRNAポリメラーゼの修飾分子基盤 (2013)
  • Author(s): 荻朋男
  • Organizer: 2013年度国立遺伝学研究所研究会「染色体DNAの安定維持の分子メカニズム」
  • Place of Presentation: 国立遺伝学研究所（三島市）
• [Presentation] DNA repair deficient genetic disorders and their clinical diagnosis (2013)
  • Author(s): 荻朋男
  • Organizer: CS meeting Amy and freiends
  • Place of Presentation: St David's Park Hotel（イギリス）
• [Presentation] 紫外線高感受性症候群～UVSSA遺伝子の発見と病態解析 (2013)
  • Author(s): 荻朋男
  • Organizer: 日本美容皮膚科学会・学術大会
  • Place of Presentation: 神戸国際会議場（神戸市）
  • Invited
• [Presentation] DNA 修復欠損性遺伝性疾患の責任因子の検索と分子機構の解析 (2013)
  • Author(s): 荻朋男
  • Organizer: 日本環境変異原学会-変異機構研究会 第26回夏の学校
  • Place of Presentation: アイプラザ一宮（一宮市）
  • Invited
• [Presentation] Repair of UV-induced DNA damage (2013)
  • Author(s): 荻朋男
  • Organizer: Youth Scientific School "Problems of Basic and Applied Radiobiology"
  • Place of Presentation: Medical Radiological Research Center（ロシア）
• [Presentation] Mutations in UVSSA cause UV-sensitive syndrome and impair RNA polymerase IIo processing in transcription-coupled nucleotide-excision repair (2012)
  • Author(s): 荻 朋男
  • Organizer: 3R meeting
  • Place of Presentation: 兵庫
• [Presentation] Molecular cloning and characterisation of KIAA1530/UVSSA, the responsible gene for UV sensitive syndrome complementation group-A (2012)
  • Author(s): 荻 朋男
  • Organizer: BIO JAPAN
  • Place of Presentation: 横浜
  • Invited
• [Presentation] 紫外線高感受性症候群責任因子UVSSAによる損傷停止RNAポリメラーゼのユビキチン化修飾メカニズムの解析 (2012)
  • Author(s): 荻 朋男
  • Organizer: 日本遺伝学会第84回大会
  • Place of Presentation: 福岡
• [Presentation] Molecular cloning and characterisation of KIAA1530/UVSSA, the responsible gene for UV sensitive syndrome complementation group-A (2012)
  • Author(s): 荻 朋男
  • Organizer: 駿河台国際シンポジウム
  • Place of Presentation: 東京
  • Invited
• [Presentation] ヒトDNA修復遺伝子の同定と機能解析
  • Author(s): 荻朋男
  • Organizer: The International Kick-Off Symposium for Atomic Bomb Disease Institute at Nagasaki University
  • Place of Presentation: 長崎大学医学部 良順会館（長崎市）
  • Invited
• [Presentation] 紫外線感受性症候群責任遺伝子UVSSAの同定と分子機能解析
  • Author(s): 荻 朋男
  • Organizer: 日本皮膚科学会
  • Place of Presentation: 長崎
  • Invited
• [Remarks] 長崎大学 がん・ゲノム不安定性研究拠点ホームページ
  • URL: http://www.nrgic.prj.nagasaki-u.ac.jp/
• [Remarks] 長崎大学がん・ゲノム不安定性研究拠点ホームページ
  • URL: http://www.nrgic.prj.nagasaki-u.ac.jp/
• [Remarks] 長崎大学 がん・ゲノム不安定性研究拠点ホームページ
  • URL: http://www.nrgic.prj.nagasaki-u.ac.jp/