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1996 Fiscal Year Final Research Report Summary

Function and sorting of yeast peroxisomal membrane proteins

Research Project

Project/Area Number 07044196
Research Category

Grant-in-Aid for international Scientific Research

Allocation TypeSingle-year Grants
SectionJoint Research
Research InstitutionKYOTO UNIVERSITY

Principal Investigator

KATO Nobuo  Faculty of Agriculture, Kyoto Univ., 農学部, 教授 (50026556)

Co-Investigator(Kenkyū-buntansha) YURIMOTO Hiroya  Faculty of Agriculture, Kyoto Univ., 農学部, 助手 (00283648)
グッドマン ジョエル・M  テキサス大学, サウスウエスタン医学センター, 助教授
GOODMAN Joel M  Southwestern Medcal Center, Texas Univ.
Project Period (FY) 1995 – 1996
KeywordsCandida boidinii / Saccharomyces cerevisiae / peroxisome / Pmp30 / Pmp27 / Pmp47 / dihydroxyacetone synthase
Research Abstract

In this project, we analyzed the relation between the biochemical functions of peroxisomal membrane proteins (PMP) and the assembly of peroxisomes, and the sorting signal for PMP,dealing with the methlotrophic yeast Candida boidinii as a model organism.
By the analysis of regulation of the expression of peroxisomal matrix and membrane proteins, we conclude that the induction of PMP occurs before the matrix enzymes and that this is regulated mainly at the mRNA level. We cloned the PMP20, PMP30, PMP47 gene from C.boidinii, disrupted these genes, and analyzed growing activity of the disruption strains (pmp30DELTA and pmp47DELTA) on several carbon sources. Both strains could grow on oleate and D-alanine, but not on methanol ; these growth substrates require peroxisomal metabolism.
The results from the electron microscopic observation of pmp30DELTA showed that Pmp30 directly promotes peroxisome proliferation and affects organelle size and shape. And also we confirmed that Pmp30 is homologous … More to Pmp27 from Saccharomyces cerevisiae by genetic complementation experiments.
Methanol-induced cells of pmp47DELTA were investicated in detail. The activity of dihydroxyacetone synthase (DHAS), which is a methanol-induced peroxisome matrix enzyme, was not detected in pmp47DELTA. Further biochemical and immunocytochemical experiments revealed that the DHAS protein aggregated in the cytoplasm as an inclusion body. Then, the PMP47 gene of two peroxisome-deficient mutants were disrupted. In these mutant strains, DHAS was enzymatically active and found in the cytoplasm. So we propose a mechanism that an unknown small molcule, which transports by Pmp47, is necessary for the folding or the translocation machinery of DHAS within peroxisomes.
Next, we analyzed the targeting sequence for peroxisomal integral membrane protein Pmp47, which spans the membrane six times. We demonstrated that the 20 amino acid loop, which predicted to face the matrix, is necessary and sufficient for sorting to peroxisomes. This is the first report of a sorting sequence, termed mPTS,for a peroxisomal integral membrane protein. Less

  • Research Products

    (12 results)

All Other

All Publications (12 results)

  • [Publications] Yasuyoshi SAKAI et al.: "The Candida boidinii peroxisomal membrane protein Pmp30 has a role in peroxisomal proliferation and is functionally homologous to Pmp27 from Saccharomyces cerevisiae" Journal of Bacteriology. 177・23. 6773-6781 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yasuyoshi SAKAI et al.: "Isolation and characterization of mutants of the methylotrophic yeast,Candida boidinii S2 that are impaired in growth on peroxisome-inducing carbon sources" Bioscience,Biotechnology,and Biochemistry. 59・5. 869-875 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Pamela A.Marshall et al.: "Pmp27 promotes peroxisomal proliferation" The Journal of Cell Biology. 129・2. 345-355 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yasuyoshi SAKAI et al.: "The absence of Pmp47,a putative yeast peroxisomal transporter,causes a defect in transport and folding of specific matrix enzyme" The Journal of Cell Biology. 134・1. 37-51 (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] John M.Dyer et al.: "The sorting sequence of the peroxisomal integral membrane protein PMP47 is contained within a short hydrophilic loop" The Journal of Cell Biology. 133・2. 269-280 (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yasuyoshi SAKAI et al.: "Application of cellular functions of the methylotrophic yeast.Metabolism,gene expression and posttranslational process.Microbial Growth on C1 Compounds" M.E.Lindstorm and F.R.Tabita,Kluber Academic Publication, 9 (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yasuyoshi SAKAI et al.: "The Candida boidinii peroxisomal membrane protein Pmp30 has a role in peroxisomal proliferation and is functionally homologous to Pmp27 from Saccharomyces cerevisiae" Journal of Bacteriology. 177. 6773-6781 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yasuyoshi SAKAI et al.: "Isolation and characterization of mutants of the methylotrophic yeast, Candida boidinii S2 that are impaired in growth on peroxisome-inducing carbon sources" Bioscience, Biotechnology, and Biochemistry. 59. 869-875 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Pamela A.Marshall et al.: "Pmp27 promotes peroxisomal proliferation" The Journal of Cell Biology. 129. 345-355 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yasuyoshi SAKAI et al.: "The absence of Pmp47, a putative yeast peroxisomal transporter, causes a defect in transport and folding of specific matrix enzyme" The Journal of Cell Biology. 134. 37-51 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] John M.Dyer et al.: "The sorting sequence of the peroxisomal integral membrane protein PMP47 is contained within a short hydrophilic loop" The Journal of Cell Biology. 133. 269-280 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yasuyoshi SAKAI et al.: M.E.Lindstorm and F.R.Tabita, Kluber Academic Publication. Application of cellular functions of the methylotrophic yeast. Metabolism, gene expression and posttranslational process. Microbial Growth on C1 Compounds, 252-260 (1996)

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

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Published: 1999-03-09  

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