ATPおよびGTPの酸化体による細胞内シグナル伝達機構の解明

• Project Area: Signaling functions of reactive oxygen species
• Project/Area Number: 21117512
• Research Category: Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Kyushu University
• Principal Investigator: 土本 大介 九州大学, 生体防御医学研究所, 助教 (70363348)
• Project Period (FY): 2009 – 2010
• Project Status: Completed (Fiscal Year 2010)
• Budget Amount: ¥5,980,000 (Direct Cost: ¥4,600,000、Indirect Cost: ¥1,380,000); Fiscal Year 2010: ¥2,990,000 (Direct Cost: ¥2,300,000、Indirect Cost: ¥690,000); Fiscal Year 2009: ¥2,990,000 (Direct Cost: ¥2,300,000、Indirect Cost: ¥690,000)
• Keywords: 2-OH-ATP / 8-oxo-GTP / 細胞死 / 細胞増殖抑制 / p38MAPK / 酸化ストレス

Research Abstract

(1)ATPの酸化的脱アミノ化体であるイノシン三リン酸(ITP)結合タンパク質の網羅的探索により新たなイノシンヌクレオチド分解酵素ヒトNUDT16タンパク質を同定し、NUDT16の発現抑制によりHeLa MR細胞が増殖抑制と核ゲノムDNAの不安定化を示すことを明らかにしNucleic Acids Research誌において発表した。
(2)ITP分解酵素ITPA欠損マウス由来の初代繊維芽細胞において増殖速度の低下と染色体不安定化、ssDNA抗体に対する反応部位の増加を観察し、継代中にマウスNUDT16の発現上昇に依存してこれらの現象が消失することからイノシンヌクレオチドによる細胞障害を哺乳動物細胞ではこれら2種類の酵素により防いでいることを明らかにし、Nucleic Acids Research誌において論文発表した。
(3)NUDT16またはITPAの欠損により引き起こされるゲノム不安定性がDNA修復酵素の働きによるものであることを明らかにした(投稿準備中)
(4)ATPの酸化体2-OH-ATPを分解する活性を持つMTH1を欠損したマウス由来の繊維芽細胞を用い、ヌクレオシド型である2-OH-adenosineをこの細胞の培地に加えるとMKK3/6のリン酸化に続くp38MAPKのリン酸化と、それに依存する細胞増殖抑制および細胞死が誘導されることを明らかにした。更にその際にMKK3/6をリン酸化する上流酵素を探索するための系を開発した。
(5)8-oxo-GTPを含むヌクレオチドに結合するタンパク質として新規がん関連タンパク質PRUNE2を同定し、Journal of Molecular Neuroscience誌において論文発表した。

Research Products

• [Journal Article] Cancer-related PRUNE2 protein is associated with nucleotides and is highly expressed in mature nerve tissues. (2011)
  • Author(s): Iwama E., et al.
  • Journal Title: Journal of Molecular Neuroscience Volume: Jan 14(In press)
  • Peer Reviewed
• [Journal Article] AP-Endonuclease 2 is necessary for normal B cell development and recovery of lymphoid progenitors after chemotherapeutic challenge. (2011)
  • Author(s): Jeroen E.J., et al.
  • Journal Title: Journal of Immunology Volume: 186(4) Pages: 1943-50
  • Peer Reviewed
• [Journal Article] NUDT16 is a (deoxy)inosine diphosphatase, and its deficiency induces accumulation of single strand breaks in nuclear DNA with growth arrest (2010)
  • Author(s): Iyama T., et al.
  • Journal Title: Nucleic Acids Research Volume: 38 Pages: 4834-43
  • Peer Reviewed
• [Journal Article] NUDT16 and ITPA play a dual protective role in maintaining chromosome stability and cell growth by eliminating dIDP/IDP and dITP/ITP from nucleotide pools in mammals. (2010)
  • Author(s): Abolhassani N., et al.
  • Journal Title: Nucleic Acids Research Volume: 38 Pages: 2891-903
  • Peer Reviewed
• [Journal Article] ITPA protein, an enzyme that eliminates deaminated purine nucleoside triphosphates in cells. (2010)
  • Author(s): Sakumi K., et al.
  • Journal Title: Mutation Research Volume: 703 Pages: 43-50
  • Peer Reviewed
• [Journal Article] Programmed cell death triggered by nucleotide pool damage and its prevention by MutT homolog-1 (MTH1) with oxidized purine nucleoside triphosphatase. (2010)
  • Author(s): Nakabeppu Y., et al.
  • Journal Title: Mutation Research Volume: 703 Pages: 51-58
  • Peer Reviewed
• [Journal Article] A comprehensive screening system for damaged nucleotide-binding proteins. (2010)
  • Author(s): Tsuchimoto D., et al.
  • Journal Title: Mutation Research Volume: 703 Pages: 37-42
  • Peer Reviewed
• [Journal Article] NUDT16 is a (deoxy) inosine diphosphatase, and its deficiency induces accumulation of single strand breaks in nuclear DNA with growth arrest. (2010)
  • Author(s): Iyama T, et al.
  • Journal Title: Nucleic Acids Research (In press)
  • Peer Reviewed
• [Journal Article] NUDT16 and ITPA play a dual protective role in maintaining chromosome stability and cell growth by eliminating dIDP/IDP and dITP/ITP from nucleotide pools in mammals. (2010)
  • Author(s): Abolhassani N, et al.
  • Journal Title: Nucleic Acids Research Jan 15(Epub ahead of print)
  • Peer Reviewed
• [Journal Article] Hydrogen in drinking water reduces dopaminergic neuronal loss in the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine mouse model of Parkinson's disease. (2009)
  • Author(s): Fujita K, et al.
  • Journal Title: PLoS One 4
  • Peer Reviewed
• [Journal Article] Apex2 is required for efficient somatic hypermutation but not for class switch recombination of immunoglobulin genes. (2009)
  • Author(s): Sabouri Z, et al.
  • Journal Title: International Immunology 8 Pages: 947-55
  • Peer Reviewed
• [Journal Article] ITPase-deficient mice show growth retardation and die before weaning. (2009)
  • Author(s): Behmanesh M, et al.
  • Journal Title: Cell Death and Differentiation 10 Pages: 1315-22
  • Peer Reviewed
• [Presentation] NUDT16 is a (deoxy)inosine diphosphatase, and its deficiency induces accumulation of single-strand breaks in nuclear DNA and growth arrest (2011)
  • Author(s): Iyama T., et al.
  • Organizer: Keystone Symposia on Molecular and Cellular Biology, Genomic Instability and DNA Repair
  • Place of Presentation: Keystone Resort, Keystone, CO, USA
  • Year and Date: 2011-02-01
• [Presentation] NUDT16は(デオキシ)イノシンニリン酸分解酵素であり、その欠損は核DNAの一本鎖切断の蓄積と増殖抑制をもたらす (2010)
  • Author(s): 土本大介, 他4名
  • Organizer: BMB2010
  • Place of Presentation: 神戸ポートアイランド
  • Year and Date: 2010-12-08
• [Presentation] Human NUDT16 is a novel nucleotide pool sanitizing enzyme hydrolyzing (deoxy) inosine diphosphate to (deoxy) inosine monophosphate (2009)
  • Author(s): Tsuchimoto D, et al.
  • Organizer: 第32回日本分子生物学会年会
  • Place of Presentation: 神奈川県横浜市パシフィコ横浜
  • Year and Date: 2009-12-10
• [Presentation] An increased expression of NUDT16 with IDP/dIDP hydrolyzing activity in immortalized ITPA-null mouse embryonic fibroblasts suppressed ITPA-deficient phenotypes (2009)
  • Author(s): Abolhassani N, et al.
  • Organizer: 第32回日本分子生物学会年会
  • Place of Presentation: 神奈川県横浜市パシフィコ横浜
  • Year and Date: 2009-12-10
• [Presentation] Human NUDT16 Is a Novel Nucleotide Pool Sanitizing Enzyme Hydrolyzing (Deoxy) Inosine Diphosphate to (Deoxy) Inosine Monophosphate. (2009)
  • Author(s): Iyama T, et al.
  • Organizer: The 49th annual meeting of the American Society for Cell Biology
  • Place of Presentation: San Diego Convention Center, San Diego, CA, USA
  • Year and Date: 2009-12-07
• [Presentation] ヒトNUDT16は(デオキシ)イノシンニリン酸を(デオキシ)イノシンーリン酸に加水分解する新規ヌクレオチドプール浄化酵素である (2009)
  • Author(s): 土本大介, ら
  • Organizer: 第20回DNA複製・組換え・修復ワークショップ
  • Place of Presentation: 滋賀県彦根市琵琶湖コンファレンスセンター
  • Year and Date: 2009-11-01
• [Presentation] Identification of the full-length form of Prune2, a neuroblastoma-related protein. (2009)
  • Author(s): Daisuke Tsuchimoto, et al.
  • Organizer: 第32回日本神経科学大会
  • Place of Presentation: 愛知県名古屋市名古屋国際会議場
  • Year and Date: 2009-09-16