ｐ３２遺伝子ノックアウトによる活性酸素産生、電子伝達系、サイトカイン産生の変化

• Project Area: Signaling functions of reactive oxygen species
• Project/Area Number: 23117712
• 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: 康 東天 九州大学, 医学(系)研究科(研究院), 教授 (80214716)
• Project Period (FY): 2011-04-01 – 2013-03-31
• Project Status: Completed (Fiscal Year 2012)
• Budget Amount: ¥6,760,000 (Direct Cost: ¥5,200,000、Indirect Cost: ¥1,560,000); Fiscal Year 2012: ¥3,380,000 (Direct Cost: ¥2,600,000、Indirect Cost: ¥780,000); Fiscal Year 2011: ¥3,380,000 (Direct Cost: ¥2,600,000、Indirect Cost: ¥780,000)
• Keywords: p32 / ミトコンドリア / ノックアウトマウス / 翻訳 / リボソーム / 活性酸素 / 自然免疫 / 炎症性サイトカイン

Outline of Annual Research Achievements

（１）組織特異的p32ノックアウトマウスの作製：組織特異的ノックアウトマウス（β細胞、心筋、血管内皮、神経）を作製を試み、去年度のβ細胞および神経細胞特異的ホモノックアウトマウスの誕生に加えて、薬剤誘導性の心筋特異的ＫＯマウス作製に成功した。神経特異的KOマウスは脳において著明な空胞形成を起こし8週齢までに大半が死亡した。心筋特異的マウスでは、薬剤誘導後に心不全を起こしている。血管内皮に関しては、まだ新生児のゲノム解析を継続している段階である。
（２）MEF細胞株とリコンビナント蛋白質を用いたｐ32機能解析：樹立したｐ32ノックアウトMEF細胞株を解析し、ミトコンドリアＤＮＡでコードされるサブユニットを含む電子伝達系の複合体I,III,IVの活性の激減と電子伝達系活性低下はミトコンドリア内における翻訳低下が原因であることを明らかにした。p32はミトコンドリアリボソームの小サブユニットと大サブユニットの安定的会合形成に必要であること、p32がミトコンドリア内RNAシャペロンとして働いていることを示唆する結果を得た。
（3）MEF細胞株を用いたサイトカイン発現解析：ｐ32ノックアウトMEF細胞を用いて、各種炎症性サイトカインならびにインターフェロンのmRNA量をRT-PCR法で定量し、いくつかの因子で産生増加を認めている。細胞内シグナル伝達経路解析し、ＡＭＰＫが持続的に活性状態にあることを見出した。

Research Progress Status

24年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

24年度が最終年度であるため、記入しない。

Research Products

• [Journal Article] p32/gC1qR is indispensable for fetal development and mitochondrial translation: importance of its RNA binding ability. (2012)
  • Author(s): Yagi M, Uchiumi T, Takazaki S, Okuno B, Nomura M, Yoshida S, Kanki T, Kang D
  • Journal Title: Nuc Acid Res Volume: 19 Issue: 19 Pages: 9717-37
  • DOI: 10.1093/nar/gks774
  • Peer Reviewed
• [Journal Article] mtSSB may sequester UNG1 at mitochondrial ssDNA and delay uracil processing until the dsDNA conformation is restored (2012)
  • Author(s): Wollen Steen, K., Doseth, B., M, P. W., Akbari, M., Kang, D., Falkenberg, M., Slupphaug, G.
  • Journal Title: DNA Repair (Amst) Volume: 11 Pages: 82-91
  • Peer Reviewed
• [Journal Article] Arg901 in the AE1 C-terminal tail is involved in conformational change but not in substrate binding (2012)
  • Author(s): Takazaki, S, Abe, Y, Yamaguchi T, Yagi M, Ueda T, Kagn D, Hamasaki N
  • Journal Title: Biochem.Biophy.Acta Volume: 1819 Issue: 3 Pages: 658-665
  • DOI: 10.1016/j.bbamem.2011.11.019
  • Peer Reviewed
• [Journal Article] Effect of mitochondrial transcription factor a overexpression on motor neurons in amyotrophic lateral sclerosis model mice (2012)
  • Author(s): Morimoto, N., Miyazaki, K., Kurata, T., Ikeda, Y., Matsuura, T., Kang, D., Ide, T., Abe, K.
  • Journal Title: J Neurosci Res Volume: 90 Pages: 1200-1208
  • Peer Reviewed
• [Journal Article] Ribonucleoprotein Y-box binding protein-1 regulates mitochondrial oxidative phosphorylation (OXPHOS) protein expression after serum stimulation through binding to OXPHOS mRNA (2012)
  • Author(s): Matsumoto S, Uchiumi T, et al
  • Journal Title: Biochemical J Volume: 443 Issue: 2 Pages: 573-584
  • DOI: 10.1042/bj20111728
  • Peer Reviewed
• [Journal Article] Localization of mRNAs encoding human mitochondrial oxidative phosphorylation proteins (2012)
  • Author(s): Matsumoto S, Uchiumi T, Saito T, Yagi M, Takazaki S, Kanki T, Kang D
  • Journal Title: Mitochondrion Volume: 12(3) Issue: 3 Pages: 391-8
  • DOI: 10.1016/j.mito.2012.02.004
  • Peer Reviewed
• [Journal Article] Mitophagy plays an essential role in reducing mitochondrial production of reactive oxygen species and mutation of mitochondrial DNA by maintaining mitochondrial quantity and quality in yeast (2012)
  • Author(s): Kurihara Y, Kanki T, Aoki Y, Hirota Y, Saigusa T, Uchiumi T, Kang D
  • Journal Title: J Biol Chem Volume: 287 Issue: 5 Pages: 3265-3272
  • DOI: 10.1074/jbc.m111.280156
  • Peer Reviewed
• [Journal Article] Recombinant mitochondrial transcription factor A protein inhibits nuclear factor of activated T cells signaling and attenuates pathological hypertrophy of cardiac myocytes (2012)
  • Author(s): Fujino, T., Ide, T., Yoshida, M., Onitsuka, K., Tanaka, A., Hata, Y., Nishida, M., Takehara, T., Kanemaru, T., Kitajima, N., Takazaki, S., Kurose, H., Kang, D., Sunagawa, K.
  • Journal Title: Mitochondrion Volume: 12 Pages: 449-458
  • Peer Reviewed
• [Journal Article] Protein instability and functional defects caused by mutations of dihydro-orotate dehydrogenase in Miller syndrome patients (2012)
  • Author(s): Fang, J., Uchiumi, T., Yagi, M., Matsumoto, S., Amamoto, R., Saito, T., Takazaki, S., Kanki, T., Yamaza, H., Nonaka, K., Kang, D.
  • Journal Title: Bioscience reports Volume: 32 Pages: 631-639
  • Peer Reviewed
• [Journal Article] The role of TFAM-associated proteins in mitochondrial RNA metabolism (2012)
  • Author(s): Uchiumi, T., Kang, D.
  • Journal Title: Biochimica et biophysica acta Volume: 1820 Pages: 565-570
  • Peer Reviewed
• [Journal Article] Mitochondrial Genomic Disintegrity and Age-Related Diseases (2012)
  • Author(s): Kang, D.
  • Journal Title: Anti-Aging Medicine Volume: 9 Pages: 201-206
  • Peer Reviewed
• [Journal Article] The physiological role of mitophagy: new insights into phosphorylation events. (2012)
  • Author(s): Hirota, Y., Kang, D., Kanki, T.
  • Journal Title: International journal of cell biology Volume: 2012 Pages: 354914-354914
  • DOI: 10.1155/2012/354914
  • Peer Reviewed
• [Journal Article] The role of TFAM-associated proteins in mitochondrial RNA metabolism (2012)
  • Author(s): Uchiumi T, Kang D
  • Journal Title: Biochim Biophys Acta Volume: 1820 Issue: 5 Pages: 565-570
  • DOI: 10.1016/j.bbagen.2011.08.014
  • Peer Reviewed
• [Journal Article] mtSSB may sequester UNG1 at mitochondrial ssDNA and delayuracil processing until the dsDNA conformation is restored (2012)
  • Author(s): Wollen Steen, K., Doseth, B., M, P.W., Akbari, M., Kang, D., Falkenberg, M., Slupphaug, G.
  • Journal Title: DNA Repair (Amst) Volume: 11 Pages: 82-91
  • Peer Reviewed
• [Journal Article] Mitochondrial p32/C1QBP is highly expressed in prostate cancer and is associated with shorter PSA relapse time after radical prostatectomy (2011)
  • Author(s): Amamoto R, Yagi M, Song Y, Oda Y, Tsuneyoshi M, Naito S, Yokomizo A, Kuroiwa K, Tokunaga S, Kato S, Hiura H, Samori T, Kang D, Uchiumi T
  • Journal Title: Cancer Sci Volume: 102 Issue: 3 Pages: 639-647
  • DOI: 10.1111/j.1349-7006.2010.01828.x
  • Peer Reviewed
• [Journal Article] Phosphorylation of Serine 114 on Atg32 mediates mitophagy (2011)
  • Author(s): Aoki, Y., Kanki, T., Hirota, Y., Kurihara, Y., Saigusa, T., Uchiumi, T., Kang, D.
  • Journal Title: Mol Biol Cell Volume: 22 Issue: 17 Pages: 3206-3217
  • DOI: 10.1091/mbc.e11-02-0145
  • Peer Reviewed
• [Journal Article] Frequent Truncating Mutation of TFAM Induces Mitochondrial DNA Depletion and Apoptotic Resistance in Microsatellite-Unstable Colorectal Cancer (2011)
  • Author(s): Guo, J., Zheng, L., Liu, W., Wang, X., Wang, Z., Wang, Z., French, A.J., Kang, D., Chen, L., Thibodeau, S.N., Liu, W.
  • Journal Title: Cancer Res Volume: 71 Issue: 8 Pages: 2978-2987
  • DOI: 10.1158/0008-5472.can-10-3482
  • Peer Reviewed
• [Presentation] 臨床化学の魅力：自動化の先にあるもの（会長シンポジウム「臨床検査の再生と将来展望」） (2012)
  • Author(s): 康 東天
  • Organizer: 第59回日本臨床検査医学会学術集会
  • Place of Presentation: 京都
  • Year and Date: 2012-11-29
  • Invited
• [Presentation] 臨床検査項目標準コード（Jlac10）最新情報～今後の方向性（シンポジウム「臨床検査の医療情報」） (2012)
  • Author(s): 康 東天
  • Organizer: 第32回日本医療情報学連合大会
  • Place of Presentation: 新潟
  • Year and Date: 2012-11-15
  • Invited
• [Presentation] Rna-Binding Proteins Involved in Mitochondrial Proteins Synthesis, (2012)
  • Author(s): Kang, D.
  • Organizer: 7th International Symposium on Genomic Medicine of CCH (Joint Symposium of Taiwan Society for Mitochondrial Research and Medicine/
  • Place of Presentation: Taichung, Taiwan
  • Year and Date: 2012-10-26
  • Invited
• [Presentation] ミトコンドリアゲノムの維持とヒトの疾患：活性酸素とtfamの意義（Ｗｓ「ミトコンドリアゲノムが制御する多様な生命現象」） (2012)
  • Author(s): 康 東天
  • Organizer: 第84回日本遺伝学会大会
  • Place of Presentation: 福岡
  • Year and Date: 2012-09-24
  • Invited
• [Presentation] 老化におけるミトコンドリアとtfam（シンポジウム「身から出た錆：ミトコンドリアから長寿を科学する」） (2012)
  • Author(s): 康 東天
  • Organizer: 第12回日本抗加齢医学会総会
  • Place of Presentation: 横浜
  • Year and Date: 2012-06-22
  • Invited
• [Presentation] 検査データの現状と課題（大会企画「医療情報データベースの特性と質」） (2012)
  • Author(s): 康 東天
  • Organizer: 第16回日本医療情報学会春季学術大会
  • Place of Presentation: 函館
  • Year and Date: 2012-05-31
  • Invited
• [Presentation] 日本版センチネルプロジェクトにおける標準化の課題Jlac10（Helicsチュートリアル「医療情報を二次利用するための標準化」 (2012)
  • Author(s): 康 東天
  • Organizer: 第16回日本医療情報学会春季学術大会.
  • Place of Presentation: 函館
  • Year and Date: 2012-05-31
  • Invited
• [Presentation] ミトコンドリアゲノムと酸化ストレス：Ｔｆａｍから見る細胞機能 (2012)
  • Author(s): 康 東天
  • Organizer: 第２２回生物試料分析科学会年次学術集会
  • Place of Presentation: 福岡
  • Year and Date: 2012-05-10
  • Invited
• [Presentation] Implications of mitochondrial oxidative stress in common diseases (2012)
  • Author(s): Dongchon Kang
  • Organizer: International symposium of mitochondrial biomedicine & ASMRM Council Meeting
  • Place of Presentation: Hangzhou, China
  • Year and Date: 2012-04-08
• [Presentation] Implications of Mitochondrial Oxidative Stress in Common Diseases, (2012)
  • Author(s): Kang, D.
  • Organizer: International symposium of mitochondrial biomedicine ASMRM Council Meeting.
  • Place of Presentation: Hangzhou, China
  • Year and Date: 2012-04-07
  • Invited
• [Presentation] ミトコンドリアゲノムと酸化ストレス:TFAMから見る細胞機能 (2012)
  • Author(s): 康東天
  • Organizer: 第22回生物試料分析科学会年次学術集会
  • Place of Presentation: 福岡
• [Presentation] MayTurner症候群に関連する血栓性閉塞と良好な下腹壁静脈の副側血行を呈した先天的AT欠損の若年症例 (2011)
  • Author(s): 藤波舞、伊藤和史、高岡理恵、羽室雅夫、篠原未帆、浦田美秩代、康東天、濱崎直孝
  • Organizer: 第195回日本内科学会近畿地方会
  • Place of Presentation: 大阪
  • Year and Date: 2011-09-10
• [Presentation] Importance of Mitochondrial genome in cellular functions : Oxidative stress and TFAM (2011)
  • Author(s): D.Kang
  • Organizer: 2011Mckay International symposium On mitochondrial medicine in human health and diseases
  • Place of Presentation: Taipei, Taiwan
  • Year and Date: 2011-07-09
• [Presentation] 生化学検査標準化と共通基準範囲に向けて (2011)
  • Author(s): 康東天
  • Organizer: 日常検査の現状と将来
  • Place of Presentation: 東京
• [Presentation] Phosphorylation of ser114 on ATG32 mediates mitophagy (2011)
  • Author(s): Tomotake Kanki, Yoshimasa Aoki, Yusuke Kurihara, Tetsu Saigusa, Takeshi Uchiumi, Dongchon Kang
  • Organizer: The 8th Conference of Asian Society for Mitochondrial Research and Medicine
  • Place of Presentation: Kagoshima, Japan
• [Presentation] Recombinant TFAM attenuates pathological remodeling in cardiac myocytes via inhibiting NAFAT signaling (2011)
  • Author(s): Takeo Fujino, Tomomi Ide, Masayoshi Yoshida, Atushi Tanaka, Ken Onitsuka, Yuko Hata, Takako Takehara, Shinya Takazaki, Mothohiro Nishida, Dongchon Kang, Kenji Sunagawa
  • Organizer: The 8th Conference of Asian Society for Mitochondrial Research and Medicine
  • Place of Presentation: Kagoshima, Japan
• [Presentation] ミトコンドリアDNA量と生体ホメオスクーシスの維持に関するマウスモデルによる検討 (2011)
  • Author(s): 井手友美、康東天
  • Organizer: 第84回日本生化学会大会
  • Place of Presentation: 京都
• [Book] Importance of mitochondrial genome in multiple cellular functions : A central role of TFAM in its maintenance (2011)
  • Author(s): Kang, D.
  • Total Pages: 20
  • Publisher: Research Signpost, Kerala
• [Remarks]
  • URL: http://www.med.kyushu-u.ac.jp/cclm/