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1995 Fiscal Year Final Research Report Summary

Analysis of molecular mechanisms for suppression of Thy-1 expression in ras-transformed cells.

Research Project

Project/Area Number 06670163
Research Category

Grant-in-Aid for General Scientific Research (C)

Allocation TypeSingle-year Grants
Research Field Pathological medical chemistry
Research InstitutionGifu University

Principal Investigator

OKANO Yukio  Gifu University, Medical School, Molecular Pathobiochemistry, Professor, 医学部, 教授 (10177066)

Co-Investigator(Kenkyū-buntansha) KIMURA Masashi  Gifu University, Medical School, Molecular Pathobiochemistry, Research Associate, 医学部, 助手 (40260575)
Project Period (FY) 1994 – 1995
KeywordsCa oscillation / Thy-1 / PIG-A
Research Abstract

Two possible mechanisms for the suppression of Thy-1 expression in ras-transformed DT cells are considered, 1)early breakdown of Thy-1 protein, 2)abnormalities in glycoconjugation with Thy-1 protein. Most of anomalous expression of glyco-phosphatidylinositol (GPI)-anchored proteins are known to be due to defects of enzymes catalyzing glycoconjugation or conjugation with PI.Eight complementation groups for GPI-anchored protein biosynthesis are known till now. Two (PIG-A and -F) of these are cloned from human, and only one (PIG-A) is cloned from mouse. However, we noticed that there is a difficult point in proceeding this research project. We learned from Dr.Junji Takeda (Research Institute of microbial diseases, Osaka University) that expression cloning technique is not useful if the defective trait is recessive in disomy mammalian cells. It is necessary to check if the defect is recessive or dominant by fusion experiments of NIH3T3 and DT cells first. If the defect is dominant, the defective gene could be cloned by complementation assay, whereas expression cloning can not be applied if the defect is recessive.
We analyzed the expression of PIG-A mRNA,the molecular structure of which has been clarified in mouse, using a technique of reverse-transcriptase-PCR (RT-PCR). We designed two sets of PCR primers covering mouse PIG-A mRNA,producing 544bp and 1023bp bands. The 544bp-bands were detected in both DT and NIH3T3 cells. The 1023bp-bands were detected in DT cells alone. These results contradicts our original assumption that the defect might be in PIG-A,suggesting a defect(s) other than PIG-A gene.

  • Research Products

    (12 results)

All Other

All Publications (12 results)

  • [Publications] Y.Banno: "Thrombin-mediated phosphoinositide hydrolysis in chinese hamster ovary cells overexpressing phospholipase C-δ1." J.Biol.Chem.269. 15846-15852 (1994)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] N.Yoshimi: "Reduced expression of phospholipase C-δ,a signal-transducing enzyme,in rat colon neoplasms induced by methylazoxymethanol acetate." Mol.Carcinogenesis. 11. 192-196 (1994)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Y.Aoyama: "Involvement of protein kinase Cin bradykinin-induced intracellular calcium increase in primary cultured human keratinocytes." J.Dermatol.Sci.9. 111-116 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] K.Nagata: "Possible interaction of haemoglobin with a low Mr GTP-binding protein,ram p25." Biochem.Mol.Biol.Intl.35. 507-515 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] K.Nagata: "Identification of heterotrimeric GTP-binding proteins in human megakaryoblastic leukemia cell line,MEG-01,and their alteration during cellular differentiation." Life Sci.57. 1675-1681 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] A.Imai: "A frame-shift mutation of the androgen receptor gene in a patient with receptor-negative complete testicular feminization:comparison with a single base substitution in a receptor-reduced incomplete form." Ann.Clin.Biochem.32. 482-486 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Y.Banno: "Thrombin-mediated phosphoinositide hydrolysis in chinese hamster ovary cells overexpressing phospholipase D-delta1" J.Biol.Chem.269. 15846-15852 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] N.Yoshimi: "Reduced expression of phospholipase C-delta, a signal-transducing enzyme, in rat colon neoplasms induced by methylazoxymethanol acetate." Mol.Carcinogenesis. 11. 192-196 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Y.Aoyama: "Involvement of protein kinase C in bradykinin-induced intracellular calcium increase in primary cultured human keratinocytes." J.Dermatol.Sci.9. 111-116 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] K.Nagata: "Possible interaction of haemoglobin with a low Mr GTP-binding protein, ram p25" Biochem.Mol.Biol.Intl.35. 507-515 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] K.Nagata: "Identification of heterotrimeric GTP-binding proteins in human megakaryoblastic leukemia cell line, MEG-01, and their alteration during cellular differentiation." Life Sci.57. 1675-1681 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] A.Imai: "A frame-shift mutation of the androgen receptor gene in a patient with receptor-nagative complete testicular feminization : comparison with a single base substitution in a receptor-reduced incomplete form." Ann.Clin.Biochem.32. 482-486 (1995)

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

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Published: 1997-03-04  

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