Molecular mechanism of mammalian mRNA surveillance

2004 Fiscal Year Final Research Report Summary

• Project/Area Number: 15209013
• Research Category: Grant-in-Aid for Scientific Research (A)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: General medical chemistry
• Research Institution: Yokohama City University Graduate School of Medical Science
• Principal Investigator: OHNO Shigeo Yokohama City University Graduate School of Medical Science, Department of Molecular Biology, Professor, 大学院・医学研究科, 教授 (10142027)
• Co-Investigator(Kenkyū-buntansha): HIRAI Syu-ichi Yokohama City University Graduate School of Medical Science, Department of Molecular Biology, Associate Professor, 大学院・医学研究科, 助教授 (80228759) ; SUZUKI Atsushi Yokohama City University Graduate School of Medical Science, Department of Molecular Biology, lecturer, 大学院・医学研究科, 講師 (00264606) ; MIZUNO Keiko Yokohama City University School of Medicine, Department of Molecular Biology, Assistant Professor, 医学部, 助手 (90221803) ; AKIMOTO Kazunori Yokohama City University School of Medicine, Department of Molecular Biology, Assistant Professor, 医学部, 助手 (70285104)
• Project Period (FY): 2003 – 2004
• Keywords: nonsense mutaion / mRNA degradation / mRNA surveillance / P53 / protein kinase / PIK remated protein kinase / RNA helicase / ATPase

Research Abstract

Eukaryotes possess a system termed 'nonsense-mediated mRNA decay' (NMD) or 'mRNA surveillance', by which aberrant mRNAs with premature termination codons (PTCs) are removed from cells, thereby protecting them from accumulation of nonfunctional or potentially harmful polypeptides. Thus, Nonsense-mediated mRNA decay (NMD) is a quality control mechanism of mRNA. Proteins required for NMD (UPF1,2,3, and SMG-1, SMG-5,6,7) have been identified initially from yeast and C. elegans genetics but recent experiments on mammlas are revealing the physiological meaning and the molecular mechanism of NMD in human cells. Central player of NMD is the putative surveillance complex that recognizes PTC-containing mRNA in a manner dependent on both splicing and translation. Components of the surveillance complex include UPF1, an RNA helicase, and UPF2 and UPF3. SMG-1 phosphorylates UPF1 and SMG and SMG-7 help dephosphorylation of UPF1 by PP2A. Importantly, both phosphorylation by SMG-1 and dephosphorylation by PP2A of UPF1 are required for NMD in human cells. Biochemical experiments revealed that phosphorylation and dephosphorylation of UPF1 by SMG-1 and PP2A, respectively, cause the remodeling of the surveillance complex. These observations supports the idea raised by a series of molecular genetic experiments that SMG-1-mediated phsphorylation fo UPF1 is the critical step during the recognition of PTC-containing mRNA by the surveillance complex.
Although NMD is primarily the protective mechanism, there are many cases of genetic diseases that are exacerbated by NMD. Thus, suppression of NMD might be one of the potential therapeutic approach against gene mutations that generates nonsense mRNA. We now have the tools that can modulate NMD, it is now possible to examine the contributuin of NMD in a variety of cases that involves NMD. Genetic diseases and cancer are the primary candidates for such experiments that aims to suppress NMD through inhibition of SMG-1.

Research Products

• [Journal Article] Affixin interacts with alpha-actinin and mediates integrin signaling for reorganization of F-actin induced by initial cell-substrate interaction. (2004)
  • Author(s): Yamaji S, Suzuki A, Kanamori H, Mishima W, Yoshimi R, Takasaki H, Takabayashi M, Fujimaki K, Fujisawa S, Ohno S, Ishigatsubo Y
  • Journal Title: J.Cell Biol. 165(4) Pages: 539-551
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Inhibition of nonsense-mediated mRNA decay rescues the phenotype in Ullrich's disease. (2004)
  • Author(s): Usuki F, Yamashita A, Higuchi I, Ohnishi T, Shiraishi T, Osame M, Ohno S
  • Journal Title: Ann.Neurol. 55(5) Pages: 740-744
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Role of the PAR-3-KIF3 complex in the establishment of neuronal polarity. (2004)
  • Author(s): Nishimura T, Kato K, Yamaguchi T, Fukata Y, Ohno S, Kaibuchi K
  • Journal Title: Nat.Cell Biol. 6(4) Pages: 328-334
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] The first CH domain of affixin activates Cdc42 and Rac1 through a1 phaPIX, a Cdc42/Rac1-specific guanine nucleotide exchanging factor. (2004)
  • Author(s): Mishima W, Suzuki A, Yamaji S, Yoshimi R, Ueda A, Kaneko T, Tanaka J, Miwa Y, Ohno S, Ishigatsubo Y
  • Journal Title: Genes Cells 9(3) Pages: 193-204
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] functional adhesion molecules (JAMs) : more molecules with dual functions? (2004)
  • Author(s): Ebnet K, Suzuki A, Ohno S, Vestweber D
  • Journal Title: J.Cell Sci. 117(Pt1) Pages: 19-29
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] hUPF1のリン酸化と脱リン酸化によるmRNA監視複合体リモデリング (2004)
  • Author(s): 山下暁朗, 鹿島勲, 大野茂男
  • Journal Title: 実験医学 22(4) Pages: 522-525
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Affixin interacts with α-actin and mediates integrin signaling for reorganization of F-actin induced by initial cell-substrate interaction (2004)
  • Author(s): Yamaji, S., Suzuki, A., Kanamori, H., Mishima, W., Yoshimi, R., Takasaki, H., Takabayashi, M., Fujimaki, K., Fujisawa, S., Ohno, S., Ishigatsubo, Y.
  • Journal Title: J. Cell Biology 165(4) Pages: 539-551
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Inhibition of nonsense-mediated mRNA decay rescues the phenotype in Ullrich's disease (2004)
  • Author(s): Usuki, F., Yamashita, A., Higuchi, I., Ohnishi, T., Shiraishi, T., Osame, M., Ohno, S.
  • Journal Title: Annals of Neurology 55 Pages: 740-744
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Role of the PAR-3-KIF3 complex in the establishment of neuronal polarity (2004)
  • Author(s): Nishimura, T., Kato, K., Yamaguchi, T., Fukata, Y., Ohno, S., Kaibuchi, K.
  • Journal Title: Nat. Cell Biol. 6(4) Pages: 328-334
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] The first CH domain of affixin activates Cdc42 and Rac1 through alphaPIX, a Cdc42/Rac1-specific guanine nucleotide exchanging factor (2004)
  • Author(s): Mishima, W., Suzuki, A., Yamaji, S., Yoshimi, R., Ueda, A., Kaneko, T., Tanaka, J., Miwa, Y., Ohno, S., Ishigatsubo, Y.
  • Journal Title: Genes Cells 9(3) Pages: 193-204
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Junctional adhesion molecules (JAMs): more molecules with dual functions? (2004)
  • Author(s): Ebnet, K., Suzuki, A., Ohno, S., Vestweber, D.
  • Journal Title: J. Cell Sci. 117(Pt 1) Pages: 19-29
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Phosphorylation of hUPF1 Induces Formation of mRNA Surveillance Complexes Containing hSMG-5 and hSMG-7. (2003)
  • Author(s): Ohnishi T, Yamashita A, Kashima I, Schell T, Anders K R, Grimson A, Hachiya T, Hentze M W, Anderson P, Ohno S
  • Journal Title: Molecular Cell 12(5) Pages: 1187-1200
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Self-association of PAR-3 mediated by the conserved N-terminal domain contributes to the development of epithelial tight junctions. (2003)
  • Author(s): Mizuno K, Suzuki A, Hirose T, Kitamura K, Kutsuzawa K, Futaki M, Amano Y, Ohnos S
  • Journal Title: J.Biol.Chem 278(33) Pages: 31240-31250
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Mammalian Lgl forms a protein complex with PAR-6 and aPKC independently of PAR-3 to regulate epithelial cell polarity. (2003)
  • Author(s): Yamanaka T, Horikoshi Y, Sugiyama Y, Ishiyama C, Suzuki A, Hirose T, Iwamatsu A, Shinohara A, Ohno S
  • Journal Title: Current Biology 13(9) Pages: 734-743
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] The second phase activation of protein kinase Cd at late G1 is required for DNA synthesis in serum-induced cell cycle progression. (2003)
  • Author(s): Kitamura K, Mizuno K, Etoh A, Akita Y, Miyamoto A, Nakayama K-I, Ohno S
  • Journal Title: Genes to Cells 8 Pages: 311-324
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Loss of von Hippel-Lindau protein causes cell density-dependent deregulation of CyclinD1 expression through Hypoxia-inducible factor. (2003)
  • Author(s): Baba M, Hirai S-I, Yamada-Okabe H, Hamada K, Tabuchi H, Kobayashi K, Kondo K, Yoshida M, Yamashita A, Kishida T, Nakaigawa N, Nagashima Y, Kubota Y, Yao M, Ohno S
  • Journal Title: Oncogene 22(18) Pages: 2728-2738
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Protein kinase C 1/I (PKC 1/I): aPKC isotype essential for the development of multicellular organism. (2003)
  • Author(s): Suzuki A, Akimoto K, Ohno S
  • Journal Title: J.Biochem (Tokyo) 133(1) Pages: 9-16
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Phosphorylation of hUPF1 Induces Formation of mRNA Surveillance Complexes Containing hSMG-5 and hSMG-7 (2003)
  • Author(s): Ohnishi, T., Yamashita, A., Kashima, I., Schell, T., Anders, K.R., Grimson, A., Hachiya, T., Hentze, M.W., Anderson, P., Ohno, S.
  • Journal Title: Molecular Cell 12 Pages: 1187-1200
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Self-association of PAR-3 mediated by the conserved N-terminal domain contributes to the development of epithelial tight junctions (2003)
  • Author(s): Mizuno, K., Suzuki, A., Hirose, T., Kitamura, K., Kutsuzawa, K., Futaki, M., Amano, Y., Ohno, S.
  • Journal Title: J. Biol. Chem. 278(33) Pages: 31240-31250
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Mammalian Lg1 forms a protein complex with PAR-6 and aPKC independently of PAR-3 to regulate epithelial cell polarity (2003)
  • Author(s): Yamanaka, T., Horikoshi, Y., Sugiyama, Y., Ishiyama, C., Suzuki, A., Hirose, T., Iwamatsu, A., Shinohara, A., Ohno, S.
  • Journal Title: Current Biology 13 Pages: 734-743
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] The second phase activation of protein kinase C δ at late G1 is required for DNA synthesis in serum-induced cell cycle progression (2003)
  • Author(s): Kitamura, K., Mizuno, K., Etoh, A., Akita, Y., Miyamoto, A., Nakayama, K-I., Ohno, S.
  • Journal Title: Genes to Cells 8 Pages: 311-324
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Loss of von Hippel-Lindau protein causes cell density-dependent deregulation of CyclinD1 expression through Hypoxia-inducible factor (2003)
  • Author(s): Baba, M., Hirai, S-I., Yamada-Okabe, H., Hamada, K., Tabuchi, H., Kobayashi, K., Kondo, K., Yoshida, M., Yamashita, A., Kishida, T., Nakaigawa, N., Nagashima, Y., Kubota, Y., Yao, M., Ohno, S.
  • Journal Title: Oncogene 22 Pages: 2728-2739
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Protein Kinase Cl/i (PKCl/i): A PKC isotype essential for the development of multicellular organisms (2003)
  • Author(s): Suzuki, A., Akimoto, K., Ohno, S.
  • Journal Title: J. Biochemistry 133 Pages: 9-16
  • Description: 「研究成果報告書概要(欧文)」より
• [Patent(Industrial Property Rights)] SMG-1特異的siRNA及びmRNAサーベイランス機構抑制剤 (2003)
  • Inventor(s): 大野 茂男, 山下 暁朗, 大西 哲生
  • Industrial Property Rights Holder: 科学技術振興事業団, 横浜市
  • Industrial Property Number: 特願2003-191243号
  • Filing Date: 2003-07-03
  • Description: 「研究成果報告書概要(和文)」より
• [Patent(Industrial Property Rights)] NMD阻害剤のスクリーニング方法 (2003)
  • Inventor(s): 大野 茂男, 鹿島 勲
  • Industrial Property Rights Holder: セルフリーサイエンス
  • Industrial Property Number: 特願2004-304925号
  • Filing Date: 2003-10-20
  • Description: 「研究成果報告書概要(和文)」より