ミトコンドリア蛋白質の翻訳異常に伴う活性酸素の産生と細胞周期連関

• Project Area: Signaling functions of reactive oxygen species
• Project/Area Number: 21117511
• 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, 医学研究院, 教授 (80214716)
• Co-Investigator(Kenkyū-buntansha): 内海 健 九州大学, 大学院・医学研究院, 准教授 (80253798) ; 神吉 智丈 九州大学, 大学病院, 助教 (50398088) ; 高崎 伸也 九州大学, 大学病院, 助教 (90435149)
• Project Period (FY): 2009 – 2010
• Project Status: Completed (Fiscal Year 2010)
• Budget Amount: ¥6,760,000 (Direct Cost: ¥5,200,000、Indirect Cost: ¥1,560,000); Fiscal Year 2010: ¥3,380,000 (Direct Cost: ¥2,600,000、Indirect Cost: ¥780,000); Fiscal Year 2009: ¥3,380,000 (Direct Cost: ¥2,600,000、Indirect Cost: ¥780,000)
• Keywords: ミトコンドリアDNA / 遺伝子診断 / 活性酸素 / リボソーム / ミトコンドリア病 / 翻訳

Research Abstract

ミトコンドリアには独自のゲノムがあり、電子伝達系に関わる蛋白を13コードしている。ミトコンドリアDNA維持機構は厳重に調整されていることが考えられているがその分子機構は明らかでない。我々はミトコンドリアDNA維持に関るTFAMに注目しTFAMに結合する蛋白を免疫沈降法で多数同定してきた。このうちERAL-1に着目しその機能解析を行った。本研究ではミトコンドリアDNA維持と翻訳に関わるERAL1およびp32蛋白質の機能について研究した。(方法と結果)(1)ERAL1に結合するRNA、蛋白を免疫沈降法で確認したところ、ERAL1はミトコンドリア12S rRNAと結合することを見出し、さらにミトコンドリア翻訳因子と多数結合することを見出した。sucrose gradientによるERAL1の局在はリボソーム小サブユニットと共局在していた。ERAL1 siRNAをHeLa細胞に処理したところミトコンドリア膜電位の低下を観察した。さらにROSの産生増加を確認した。ERAL1ノックダウンにより細胞はG2/M arrest, apoptosisを起こすことを見出した。これらの結果はERAL1はミトコンドリアリボソーム小サブユニットに結合し翻訳を制御することを示唆している。(2)p32の全身ノックアウトマウスを作製したところ、胎生10週前後で致死であった。この胎児マウスからMEF細胞株を樹立し、ミトコンドリア機能を測定したところ、電子伝達系活性が激減し、おそらくそれに付随してと思われるROSの産生増加を見出した。活性低下はmtDNAコード蛋白質の翻訳障害が原因であった。現在の所、p32はミトコンドリアリボソームにmRNAを結合させ翻訳を開始させる経路に必要であることを示唆する結果を得ている。

Research Products

• [Journal Article] Structure of the membrane domain of human erythrocyte anion exchanger 1 revealed by electron crystallography. (2010)
  • Author(s): Yamaguchi, T., Ikeda, Y., Abe, Y., Kuma, H., Kang, D., Hamasaki, N., Hirai, T.
  • Journal Title: J Mol Biol Volume: 397 Pages: 179-189
  • Peer Reviewed
• [Journal Article] Helical image reconstruction of the outward-open human erythrocyte band 3 membrane domain in tubular crystals (2010)
  • Author(s): Yamaguchi, T., Fujii, T., Abe, Y., Hirai, T., Kang, D., Namba, K., Hamasaki, N., Mitsuoka, K.
  • Journal Title: J Struct Biol Volume: 169 Pages: 406-412
  • Peer Reviewed
• [Journal Article] ERAL1 is associated with mitochondrial ribosome and elimination of ERAL1 leads to mitochondrial dysfunction and growth retardation (2010)
  • Author(s): Uchiumi, T., Ohgaki, K., Yagi, M., Aoki, Y., Sakai, A., Matsumoto, S., Kang, D
  • Journal Title: Nucleic Acids Res Volume: 38 Pages: 5554-5568
  • NAID: 120005227389
  • Peer Reviewed
• [Journal Article] Geographic distribution of fluoroquinolone-resistant Escherichia coli strains in Asia (2010)
  • Author(s): Uchida, Y., Mochimaru, T., Morokuma, Y., Kiyosuke, M., Fujise, M., Eto, F., Harada, Y., Kadowaki, M., Shimono, N., Kang, D.
  • Journal Title: Int J Antimicrob Agents Volume: 35 Pages: 387-391
  • Peer Reviewed
• [Journal Article] Clonal spread in Eastern Asia of ciprofloxacin-resistant Escherichia coli serogroup O25 strains, and associated virulence factors. Int (2010)
  • Author(s): Uchida, Y., Mochimaru, T., Morokuma, Y., Kiyosuke, M., Fujise, M., Eto, F., Eriguchi, Y., Nagasaki, Y., Shimono, N., Kang, D.
  • Journal Title: J Antimicrob Agents Volume: 35 Pages: 444-450
  • Peer Reviewed
• [Journal Article] Mutation of His 834 in human anion exchanger 1 affects substrate binding (2010)
  • Author(s): Takazaki, S., Abe, Y., Yamaguchi, T., Yagi, M., Ueda, T., Kang, D., Hamasaki, N
  • Journal Title: Biochim Biophys Acta Volume: 1798 Pages: 903-908
  • Peer Reviewed
• [Journal Article] PLEDs in an infant with congenital protein C deficiency : a case report (2010)
  • Author(s): Sekiguchi, K., Akiyoshi, K., Okazaki, N., Yamada, H., Suzuki, M., Maeda, T., Suenobu S., Izumi, T., Kang, D.
  • Journal Title: Clin Neurophysiol Volume: 121 Pages: 800-801
  • Peer Reviewed
• [Journal Article] IFCC reference procedures for measurement of the catalytic concentrations of enzymes : corrigendum, notes and useful advice (2010)
  • Author(s): Schumann, G., Canalias, F., Joergensen, P.J., Kang, D., Lessinger, J.M., Klauke, R
  • Journal Title: Clin Chem Lab Med Volume: 48 Pages: 615-621
  • Peer Reviewed
• [Journal Article] Mitochondrial single-stranded DNA binding protein is required for maintenance of mitochondrial DNA and 7S DNA but is not required for mitochondrial nucleoid organisation (2010)
  • Author(s): Ruhanen, H., Borrie, S., Szabadkai, G., Tyynismaa, H., Jones, A.W., Kang, D., Taanman, J.W., Yasukawa, T
  • Journal Title: Biochim Biophys Acta Volume: 1803 Pages: 931-939
  • Peer Reviewed
• [Journal Article] Mutation of His 834 in human anion exchanger 1 affects substrate binding. Biochim (2010)
  • Author(s): Takazaki, S., et al.
  • Journal Title: Biochim Biophys Acta 1798 Pages: 903-908
  • Peer Reviewed
• [Journal Article] Helical image reconstruction of the outward-open human erythrocyte band 3 membrane domain in tubular crystals. (2010)
  • Author(s): Yamaguchi, T., et al.
  • Journal Title: J Struct Biol 169 Pages: 406-412
  • Peer Reviewed
• [Journal Article] Structure of the membrane domain of human erythrocyte anion exchanger 1 revealed by electron crystallography. (2010)
  • Author(s): Yamaguchi, T., et al.
  • Journal Title: J Mol Biol 397 Pages: 179-189
  • Peer Reviewed
• [Journal Article] DNA conformation-dependent activities of human mitochondrial RNA polymerase. (2009)
  • Author(s): Fukuoh, A., et al.
  • Journal Title: Genes Cells 14 Pages: 1029-1042
  • Peer Reviewed
• [Journal Article] Development of a new measurement method for serum calcium with chlorophosphonazo-III. (2009)
  • Author(s): Hokazono, E., et al.
  • Journal Title: Ann Clin Biochem 46 Pages: 296-301
  • Peer Reviewed
• [Journal Article] Fulminant sepsis/meningitis due to Haemophilus influenzae in a protein C-deficient heterozygote treated with activated protein C therapy. (2009)
  • Author(s): Ishimura, M., et al.
  • Journal Title: Eur J Pediatr 168 Pages: 673-677
  • Peer Reviewed
• [Journal Article] High-dose penicillin G-treatment causes underestimation of serum albumin measured by a modified BCP method. (2009)
  • Author(s): Ono, M., et al.
  • Journal Title: Clin. Chim. Acta 407 Pages: 75-76
  • Peer Reviewed
• [Journal Article] Human heart mitochondrial DNA is organized in complex catenated networks containing abundant four-way junctions and replication forks. (2009)
  • Author(s): Pohjoismaki, J.L., et al.
  • Journal Title: J Biol Chem 284 Pages: 21446-21457
  • Peer Reviewed
• [Journal Article] Association of a novel mitochondrial protein M19 with mitochondrial nucleoids. (2009)
  • Author(s): Sumitani, M., et al.
  • Journal Title: J Biochem 146 Pages: 725-732
  • Peer Reviewed
• [Presentation] Aging and mitochondrial dysfunction (2010)
  • Author(s): Kang, D.
  • Organizer: The 11th annual meeting for Southern district of the Korean Society for Laboratory Medicine
  • Place of Presentation: Pusan, Korea
• [Presentation] TFAM-associated proteins that are involved in mitochondrial RNA metabolisms (2010)
  • Author(s): Kang, D
  • Organizer: The 7th Conference of Asian Society for Mitochondrial Research and Medicine
  • Place of Presentation: Fukuoka, Japan
• [Remarks]
  • URL: http://www.med.kyushu-u.ac.jp/cclm/kyositu/index.html