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2003 Fiscal Year Final Research Report Summary

Molecular genetic studies of control of mRNA stability in the gene for the key enzyme of methionine biosynthesis

Research Project

Project/Area Number 13440233
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field 植物生理
Research InstitutionHokkaido University

Principal Investigator

NAITO Satoshi  Hokkaido Univ., Grad. School of Agr., Prof., 大学院・農学研究科, 教授 (20164105)

Co-Investigator(Kenkyū-buntansha) ONOUCHI Hitoshi  Hokkaido Univ., Grad. School of Agr., Inst., 大学院・農学研究科, 助手 (50322839)
Project Period (FY) 2001 – 2003
KeywordsArabidopsis / mutant / methionine / in vitro translation / mRNA stability / cystathionine gamma-synthase
Research Abstract

Expression of the gene for cystathionine gamma-synthase (CGS), the key enzyme of methionine biosynthesis in higher plants is feedback-regulated at the step of mRNA stability and the amino acid sequence in the exon 1 of CGS itself is involved in this regulation. In order to identity the factors that are involved in this regulation, a chimeric gene carrying green fluorescent protein gene (GFP) fused to the exon 1 of CGS gene and placed under the control of a cauliflower mosaic virus 35S promoter was constructed and introduced into wild-type Arabidopsis. The transgenic line termed GFPc4 was used to screen for those mutants that show higher GFP fluorescence relative to the autofluorescence of chloroplasts. Four of the mutants thus isolated seemed to affect the expression of CGS in traps. One of them showed elevated levels of both mRNAs for endogenous CGS and exogenous GFP fusion genes and reduced down-regulation of stability of mRNAs of both endogenous CGS and exogenous GFP fusion genes in response to methionine application. Transient expression analyses by electroporation to the mutant protoplasts were carried out A similar chimeric gene was constructed using E coli beta-glucuronidase gene as a reporter and the effects of methionine and S-adenosylmethionine (SAM) in the culture medium were tested. The results showed that in the mutant, the effects of both methionine and SAM were reduced as compared to the parental line. SAM has been shown to be the efftector of the post-transcriptional regulation of CGS expression. The results suggest that a mutant that affects the post-transcriptional regulation of CGS expression in trans.

  • Research Products

    (10 results)

All Other

All Publications (10 results)

  • [Publications] Suzuki, A.: "The first exon coding region of cystathionine γ-synthase gene is necessary and sufficient for downregulation of its own mRNA accumulation in transgenic Arabidopsis thaliana."Plant Cell Physiol.. 42. 1174-1180 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Goto, D.B.: "A single-nucleotide mutation in a gene encoding S-adenosylmethionine synthetase is associated with methionine over-accumulation phenotype in Arabidopsis thaliana."Genes Genet.Syst.. 77. 89-95 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Ominato, K.: "Identification of a short highly conserved amino acid sequence as the functional region required for posttranscriptional autoregulation of the cystathionine γ-synthase gene in Arabidopsis."J.Biol.Chem.. 277. 36380-36386 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Chiba, Y.: "S-Adenosyl-L-methionine is an effector in the posttranscriptional autoregulation of the cystathionine γ-synthase gene in Arabidopsis."Proc.Natl.Acad.Sci.USA. 100. 10225-10230 (2003)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Lambein, I.: "Decay kinetics of autogenously regulated CGS1 mRNA that codes for cystathionine γ-synthase in Arabidopsis thaliana."Plant Cell Physiol.. 44. 893-900 (2003)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Suzuki, A.: "The first exon coding region of cystathionine γ-synthase gene is necessary and sufficient for downregulation of its own mRNA accumulation in transgenic Arabidopsis thaliana."Plant Cell Physiol.. 42. 1174-1180 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Goto, D.B.: "A single-nucleotide mutation in a gene encoding S-adenosylmethionine synthetase is associated with methionine over-accumulation phenotype in Arabidopsis thaliana."Genes Genet.Syst.. 77. 89-95 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ominato, K.: "Identification of a short highly conserved amino acid sequence as the functional region required for posttranscriptional autoregulation of the cystathionine γ-synthase gene in Arabidopsis."J.Biol.Chem.. 277. 36380-36386 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Chiba, Y.: "S-Adenosyl-L-methionine is an effector in the posttranscriptional autoregulation of the cystathionine γ-synthase gene in Arabidopsis"Proc.Natl.Acad.Sci.USA. 100. 10225-10230 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Lambein, I.: "Decay kinetics of autogenously regulated CGSI mRNA that codes for cystathionine γ-synthase in Arabidopsis thaliana."Plant Cell Physiol.. 44. 893-900 (2003)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 2005-04-19  

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