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1998 Fiscal Year Final Research Report Summary

Functional analysis of thyroid hormone receptor interacting protein

Research Project

Project/Area Number 09671045
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field 内分泌・代謝学
Research InstitutionNagoya University

Principal Investigator

NAGAYA Takashi  Res.Inst.Environ.Med., Nagoya Univ., Research Associate, 環境医学研究所, 助手 (80262913)

Co-Investigator(Kenkyū-buntansha) KAMBE Fukushi  Res.Inst.Environ.Med., Nagoya Univ., Associate Prodessor, 環境医学研究所, 助教授 (00211871)
MURATA Yoshiharu  Res.Inst.Environ.Med., Nagoya Univ., Professor, 環境医学研究所, 教授 (80174308)
SEO Hisao  Res.Inst.Environ.Med., Nagoya Univ., Professor, 環境医学研究所, 教授 (40135380)
Project Period (FY) 1997 – 1998
KeywordsThyroid hormone receptor / TR interacting protein / Yeast two hybrid system / Co-repressor / Thyroid hormone / Cloning
Research Abstract

To understand the molecular aspect of thyroid hormone action, we performed to clone TR interacting protein by yeast two hybrid system. Hela cell cDNA library constructed in Gal4 activation domain vector (pGAD-GH) was screened by human thyroid hormone receptor beta (TRbeta) in Gal4 DNA binding domain vector (pGBT9). One done (named B1) to interact with TRbeta was isolated and considered to be a partial fragment of human nuclear receptor co-repressor (hN-CoR) by nucleotide sequencing. By colony hybridization technique and 5'-RACE, we cloned wild type and other three variants of hN-CoRs. One variant (hN-CoRv1) is 56 amino acids (a.a.) insertion in to a.a. 1018 of wild type N-CoR.The second one (hN-CoRv2) is a deletion of 120 a.a between a.a. 1857 and 1977. The third one is a carboxyterminal variant, in that 120 a.a. of wild type sequence is replaced with distinct 50 a.a..
To study the chromosomal localization of hN-CoR, fluorescence in situ hybridization technique was performed using hN-Co … More R cDNA as a probe. Then the locus of hN-CoR was assigned to chromosome 17p11.2. Since hN-CoR localizes at one locus, the variants are considered to be generated by alternative splicings.
Since mN-CoR interacts with TR and retinoic acid receptor (RAR) through its carboxy-terminal region, the alteration of carboxy-terminus in N-CoR variant (clone H3) might affect specificity of receptor interactions. The existence of N-CoR isoforms will support the possibility that the different expression of these isoforms may modulate transcriptional regulation by thyroid hormone.
Expression profile of N-CoR variants was analyzed in 8 human cell lines by RT-PCR method. The extracted RNA was reverse-transcribed and followed by the amplification of N-CoR variants with PCR.In all cell lines studied, mRNA expression of two N-CoR variants (hN-CoRv2 and v3) were detected minimally in comparison with Wild type. However, the expression of hN-CoRv1 was different in the cell lines. In three cell lines (NB-1 neuroblastoma cell line, Hela cervival carcinoma cell line and HOS osteosarcoma cell line), hN-CoRv1 was expressed predominant than wild type. The diffrent expression of hN-CoR variants might modify repression level by co-repressor and modulate hormonal signalings. Less

  • Research Products

    (12 results)

All Other

All Publications (12 results)

  • [Publications] Nagaya T,他: "Localization of the human nuclear co-repressor gene between the CMT1A and SMS critical regions of chromosome 17p11.2" Genomics. in press. (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Nagaya T,他: "Intracellular proteolytic cleavage of RXRα by cathepsin L-type protease is a potential mechanism for modulating thyroid horomone action." J.Biol.Chem.273. 33166-33173 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Nagaya T,他: "Molecular basis of resistance to thyroid hormone; Review." Endocrine J.45. 709-718 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Nagaya T,他: "Requirement of corepressor binding of thyroid hormone receptor mutants for dominant negative inhibition." Biochem.Biophys.Res.Commun.247. 620-623 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Liu Y,他: "An inhibitory region of the DNA-binding domain of thyroid hormone receptor blocks hormone-dependent transactivation." Mol.Endocrinol.12. 34-44 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Tagami T,他: "Nuclear receptor corepressors activate rather than suppress basal transcription of genes that are negatively regulated by thyroid hormone." Mol.Cell.Biol.17. 2642-2648 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Nagaya T,et al.: "Localization of the human nuclear co-repressor gene between the CMT1A and SMS critical regions of chromosome 17p11.2" Genomics. (in Press). (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Nagaya T,et al.: "Intracellular proteolytic cleavage of RXRalpha by cathepsin L-type protease is a potential mechanism for modulating thyroid horomone action." J.Biol.Chem.273. 33166-33173 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Nagaya T,et al.: "Moleclular basis of resistance to thyroid homone ; Review." Endocrine J.45. 709-718 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Nagaya T,et al.: "Requirement of corepressor binding of thyroid hormone receptor mutants for dominant negative inhibition." Biochem.Biophys.Res.Commun.247. 620-623 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Liu Y,et al.: "An inhibitory region of the DNA-binding domain of thyroid hormone receptor blocks hormone-dependent transactivation." Mol.Endocrinol.12. 34-44 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Tagami T,et al.: "Nuclear receptor corepressors activate rather than suppress basal transcription of genes that are negatively regulated by thyroid hormone." Mol.Cell.Biol.17. 2642-2648 (1997)

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

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Published: 1999-12-08  

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