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Molecular mechanism of mammalian mRNA surveillance

Research Project

Project/Area Number 17209010
Research Category

Grant-in-Aid for Scientific Research (A)

Allocation TypeSingle-year Grants
Section一般
Research Field General medical chemistry
Research InstitutionYokohama City University

Principal Investigator

OHNO Shigeo  Yokohama City University, Graduate School of Medical Science, Department of Molecular Biology, Professor, 医学研究科, 教授 (10142027)

Co-Investigator(Kenkyū-buntansha) HIRAI Syu-ichi  Yokohama City thiiversity Graduate School of Medical Science, Department of Molecular Biology, Associate Professor, 医学研究科, 準教授 (80228759)
SUZUKI Atsushi  Yokohama City University Graduate School of Medical Science, Department of Molecular Biology, Associate Professor, 医学研究科, 準教授 (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) 2005 – 2006
Project Status Completed (Fiscal Year 2006)
Budget Amount *help
¥51,350,000 (Direct Cost: ¥39,500,000、Indirect Cost: ¥11,850,000)
Fiscal Year 2006: ¥24,570,000 (Direct Cost: ¥18,900,000、Indirect Cost: ¥5,670,000)
Fiscal Year 2005: ¥26,780,000 (Direct Cost: ¥20,600,000、Indirect Cost: ¥6,180,000)
Keywordsnonsense mutation / mRNA degradation / mRNA surveillance / P53 / protein kinase / PIK remated protein kinase / RNA helicase / ATPase
Research Abstract

Nonsense-mediated mRNA decay (NMD) is a surveillance mechanism that degrades mRNA containing premature termination codons (PTCs). In mammalian cells, recognition of PTCs requires translation and depends on the presence on the mRNA with the splicing-dependent exon junction complex (EJC). While it is known that a key event in the triggering of NMD is phosphorylation of the trans-acting factor, Upf1, by SMG-1, the relationship between Upf1 phosphorylation and PTC recognition remains undetermined. Here we show that SMG-1 binds to the mRNA-associated components of the EJC, Upf2, Upf3b, elF4A3, Magoh, and Y14. Further, we describe a novel complex that contains the NMD factors SMG-1 and Upf1, and the translation termination release factors eRF1 and eRF3 (SURF). Importantly, an association between SURF and the EJC is required for SMG-1-mediated Upf1 phosphorylation and NMD. Thus, the SMG-1-mediated phosphorylation of Upf1 occurs on the association of SURF with EJC, which provides the link betw … More een the EJC and recognition of PTCs and triggers NMD.
Phosphatidylinositol 3-kinase-related kinases (PIKKs) consisting of SMG-1, ATM, ATR, DNA-PKcs, and mTOR are a family of proteins involved in the surveillance of gene expression ineukaryotic cells. They are involved in mechanisms responsible for genome stability, mRNA quality, and translation. They share a large N-terminal domain and a C-terminal FATC domain in addition tothe unique serine/threonine protein kinase (PIKK) domain that is different from classical proteinkinases. However, structure-function relationships of PIKKs remain unclear. Here we have focused onone of the PIKK members, SMG-1 that is involved in RNA surveillance termed nonsense-mediatedmRNA decay (N MD), to analyze the roles of conserved and SMG-1 specific sequences on the intrinsickinase activity. Analyses of sets of point and deletion mutants of SMG-1 in a purified system and intactcells revealed that the long N-terminal region and the conserved leucine in the FATC domain wereessential for SMG-1 kinase activity. However, the conserved tryptophan in the TS domain and theFATC domain was not. In addition, the long insertion region between PIKK and FATC domains wasnot essential for SMG-1 kinase activity. These results indicated an unexpected feature of SMG-1, i.e.,the distantly located N-terminal and C-terminal sequences were essential for the intrinsic kinaseactivity.
In order to confirm our previous results using SMG-1 inhibitors, we evaluated the effects of NMD inhibition by siRNA-mediated knockdown of SMG-1 or Upf1 on the phenotype of Ullrich disease, an autosomal recessive congenital muscular dystrophy. The patient studied showed a homozygousframeshift mutation with a PTC in the collagen VI A2 gene, which encodes a truncated but partiallyfunctional protein. The patient's fibroblasts showed a nearly complete loss of the triple-helical collagenVI protein and functional defects in the extracellular matrix (ECM) due to the crucial deficiency of the collagen VI a2 protein. We have shown that siRNA-mediated knockdown of SMG-1 or Upf1 causes theupregulation of the mutant triple-helical collagen VI, resulting in the formation of partially functionalECM. We suggest that the inhibition of NMD may be useful as a therapeutic approach to treat somehuman genetic diseases exacerbated by NMD. Less

Report

(3 results)
  • 2006 Annual Research Report   Final Research Report Summary
  • 2005 Annual Research Report
  • Research Products

    (17 results)

All 2007 2006 2005 Other

All Journal Article (17 results)

  • [Journal Article] Distant N-and C-terminal domains are required for intrinsic kinase activity of SMG-1, a critical component of nonsense-mediated mRNA decay.2007

    • Author(s)
      Morita T, Yamashita A, Kashima I, Ogata K, Ishiura S, Ohno S
    • Journal Title

      J. Biol., Chem. 282(11)

      Pages: 7799-7808

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Distant N-and C-terminal domains are required for intrinsic kinase activity of SMG-1, a critical component of nonsense-mediated mRNA decay.2007

    • Author(s)
      Morita T, Yamashita A, Kashima I, Ogata K, Ishiura S, Ohno S
    • Journal Title

      J.Biol., Chem. 282(11)

      Pages: 7799-7808

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Distant N- and C-terminal domains are required for intrinsic kinase activity of SMG-1,a critical component of nonsense-mediated mRNA decay.2007

    • Author(s)
      Morita T, Yamashita A, Kashima I, 0gata K, Ishiura S, Ohno S
    • Journal Title

      J.Biol.,Chem. 282(11),

      Pages: 7799-7808

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Specific inhibition of nonsense-mediated mRNA decay components, SMG-1 or Upf1, rescues the phenotype of Ullrich's disease fibroblasts.2006

    • Author(s)
      Usuki F, Yamashita A, Kashima I, Higuchi I, Mitsuhiro, Osame M, Ohno S
    • Journal Title

      Molecular Therapy 14(3)

      Pages: 351-360

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Binding of a novel SMG-1-Upf1-eRF1-eRF3 complex (SURF) to the exon junction complex triggers Upf1phosphorylation and nonsense-mediated mRNA decay.2006

    • Author(s)
      Kashima I, Yamashita A, Izumi N, Kataoka N, Morishita R, Hoshino S, Ohno M, Dreyfuss G, Ohno S
    • Journal Title

      Genes Dev. 20(3)

      Pages: 355-367

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Annual Research Report 2006 Final Research Report Summary
  • [Journal Article] The role of SMG-1-mediated phosphorylation of UPF1 in NMD.2006

    • Author(s)
      Yamashita A, Kashima I, Ohno S
    • Journal Title

      In Nonsense-Mediated mRNA decay

      Pages: 85-95

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Annual Research Report 2006 Final Research Report Summary
  • [Journal Article] mRNA品質管理システムにおけるナンセンスコドン認識の分子機構2006

    • Author(s)
      鹿島 勲, 山下暁朗, 大野茂男
    • Journal Title

      実験医学 24(7)

      Pages: 963-966

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Specific inhibition of nonsense-mediated mRNA decay components, SMG-1 or Upfl, rescues the phenotype of Ulrich' s disease fibroblasts.2006

    • Author(s)
      Usuki F, Yamashita A, Kashima I, Higuchi I, Mitsuhiro Osame M, Ohno S
    • Journal Title

      Molecular Therapy 14, 3

      Pages: 351-360

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Binding of a novel SMG-1-Upfl-eRF1-eRF3 complex (SURF) to the exon junction complex triggers Upflphosphorylation and nonsense-mediated mRNA decay.2006

    • Author(s)
      Kashima I, Yamashita A, Izumi N, Kataoka N, Morishita R, Hoshino S, Ohno M, Dreyfuss G, Ohno S
    • Journal Title

      Genes Dev. 20(3)

      Pages: 355-367

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] The role of SMG-1-Mediated phosphorylation of UPF1 in NMD.2006

    • Author(s)
      Yamashita A, Kashima I, Ohno S
    • Journal Title

      In Nonsense-Mediated mRNA Decay, Maquat, Landes Bioscience (ISBN No. 1-58706-296-8)

      Pages: 85-95

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] Specific inhibition of nonsense-mediated mRNA decay components,SMG-1 or Upf1, rescues the phenotype of Ullrich's disease fibroblasts.2006

    • Author(s)
      Usuki F, Yamashita A, Kashima I, Higuchi I, Mitsuhiro Osame M, Ohno S
    • Journal Title

      Molecular Therapy 14,3

      Pages: 351-360

    • Related Report
      2006 Annual Research Report
  • [Journal Article] An association between the initial SMG-1-Upf1-eRF1-eRF3 complex and exon junction complex on mRNP is required for the SMG-1-mediated phosphorylation of Upf1 during nonsense-mediated mRNA decay.2006

    • Author(s)
      Kashima I, Yamashita A, Izumi N, Kataoka N, Morishita R, Hoshino S, Ohno M, Dreyfuss G, Ohno S
    • Journal Title

      Genes Dev. 20(3)

      Pages: 355-367

    • Related Report
      2005 Annual Research Report
  • [Journal Article] The role of SMG-1 on nonsense-mediated mRNA decay.2005

    • Author(s)
      Yamashita A, Kashima I, Ohno S
    • Journal Title

      Biochem Biophys. Acta (Proteins and Proteomics), Inhibitors of Protein Kinases 1754(1-2)

      Pages: 305-315

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] The role of SMG-1 on nonsense-mediated mRNA decay. Biochem Biophys. Acta (Proteins and Proteomics)2005

    • Author(s)
      Yamashita A, Kashima I, Ohno S
    • Journal Title

      Inhibitors of Protein Kinases 1754(1-2)

      Pages: 305-315

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2006 Final Research Report Summary
  • [Journal Article] The role of SMG-1 on nonsense-mediated mRNA decay.2005

    • Author(s)
      Yamashita A, Kashima I, Ohno S
    • Journal Title

      Biochem Biophys.Acta (Proteins and Proteomics), Inhibitors of Protein Kinases 1754(1-2)

      Pages: 305-315

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Role of SMG-1-Mediated Phosphorylation of Upf1 in NMD. In Nonsense-Mediated mRNA Decay,2005

    • Author(s)
      Yamashita A, Kashima I, Ohno S
    • Journal Title

      Maquat, Landes Bioscience, (ISBN No.1-58706-296-8)

      Pages: 85-95

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Specific inhibition of nonsense-mediated mRNA decay components, SMG-1 or Upf1, rescues the phenotype of Ullrich's disease fibroblasts.

    • Author(s)
      Usuki F, Yamashita A, Kashima I, Higuchi I, Mitsuhiro Osame M, Ohno S
    • Journal Title

      Molecular Therapy (in press)

    • Related Report
      2005 Annual Research Report

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Published: 2005-04-01   Modified: 2016-04-21  

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