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1999 Fiscal Year Final Research Report Summary

Functional Analysis of H1 and Core Histone Variants by Gene Targeting Technique

Research Project

Project/Area Number 09480152
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Structural biochemistry
Research InstitutionMiyazaki Medical College

Principal Investigator

NAKAYAMA Tatsuo  Miyazaki Medical College, Professor, 医学部, 教授 (60031712)

Co-Investigator(Kenkyū-buntansha) TAKECHI Shinji  Miyazaki Medical College, Assistant Professor, 医学部, 助手 (10222100)
TAKAMI Yasunari  Miyazaki Medical College, Associate Professor, 医学部, 講師 (80236356)
Project Period (FY) 1997 – 1999
KeywordsHistone variants / DT40 cells / Gene targeting technique / Chromatin structure / Transcription regulation / Histone deacetylase / IgM H-chain / Cell growth
Research Abstract

Thirty-nine of the 44 chicken H1 and core histone genes are located in a major histone gene cluster of 110 kb. Using gene targeting techniques, we generated several DT40 mutants, respectively, which are devoid of one or two particular histone genes, one allele of the major gene cluster of 110 kb, an approximately 57 kb segment of the cluster carrying 21 genes, and 11 of the 12 H1 gene copies. In addition, we generated four DT40 mutants, respectively, devoid of chHDAC-1, 2, 3 and 4. Systematic analyses of the resultant mutants led us to some noticeable conclusions, as follows.
1) Histone gene families, H1, H2A, H2B, H3 and H4, have the inherent ability to compensate for the deletion of approximately half of their own constituents and to maintain the amount of each of the histone subtypes in stoichiometric balance, based on increases in the expression of the remaining members, regardless of the complete disruption of one allele of the major gene cluster or the disruption of approximately … More half segments of the two alleles. Therefore, one allele of the major histone gene cluster is enough for cell proliferation.
2) The deletion of 11 of the 12 H1 gene copies resulted in insignificant influence on the cell functions, i.e. the growth rate and global chromatin structure, indicating that only one copy of the H1 genes is enough for cell proliferation.
3) The protein patterns on 2D-PAGE are not altered in the case of the deletion of one allele of the cluster, due to no changes in the composition of any histone variant, but vary in the case of the deletion of approximately half segments of the cluster, and the latter plus more H1 gene(s), due to changes in the quality of the H1 and core variants.
4) The protein patterns are altered in the mutants, respectively, lacking particular H1 and H2B variants. The variable proteins are specific for the corresponding mutants, suggesting that the H1 and core variants should play individual roles in regulation of the expression of specific genes, probably through alterations in the chromatin structure localized in specific genomic regions.
5) chHDAC-2 controls the amount of the IgM H-chain at the steps of both transcription of its gene and the alternative processing of its pre-mRNA.
6) chHDAC-3 is essential for the viability of DT40 cells. Less

  • Research Products

    (17 results)

All Other

All Publications (17 results)

  • [Publications] Ahmad, a. et al.: "WD repeats of the p48 subunit of chiken chromatin assembly factor-1 required for in vitro interaction with chicken"J. Biol. Chem.. 274. 16646-16653 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Takami, Y. et al.: "Chicken histone deacetylase-2 controls the amount of the IgN H-chain at the steps of both transcription of its gene and"J. Biol. Chem.. 274. 23977-23990 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Takechi, S. et al.: "Sas3 is a histone acetyltransferase and requires a zincfinger motif"Biochem. Biophys. Res. Commun.. 266. 405-410 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Nakayama, T. et al.: "Interaction of the p48 subunit of chicken chromatin assembly factor-1 with histone deacetylases"Current Topics in Biochemical Research. 1. 173-178 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Takami, Y. et al.: "Histone H1 variants play individual roles in transcription regulation in the DT40 chicken B cell line"Biochem. Biophys. Res. Commun.. (in press). (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Takami, Y. et al.: "N-terminal region, C-terminal region, nuclear export signal and deacetylation activity of histone deacetylase-3 are"J. Biol. Chem.. (in press). (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Takami, Y., Takeda, s. and Nakayama, T.: "An approximately half set of histone genes is enough for cell proliferation and a lack of several histone variants causes protein pattern changes in the DT40 chicken B cell line."J. Mol. Biol.. 265. 394-408 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Takami, Y. and Nakayama, T.: "One allele of the major histone gene cluster is enough for cell proliferation of the DT40 chicken B cell line."Biochim. Biophys. Acta. 1354. 105-115 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Takami, Y. and Nakayama, T.: "A single copy of linker H1 genes is enough for proliferation of the DT40 chicken B cell line, and linker H1 variants participate in regulation of gene expression."Genes to Cells. 2. 711-723 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Nakayama, T. and Takami, Y.: "Systematic analysis of the nature of histone genes in the chicken DT40 B cell line using gene targeting techniques."Trends in Comp. Biochem. Physiol.. 4. 211-217 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ahmad, A., Takami, Y. and Nakayama, T.: "WD repeats of the p48 subunit of chicken chromatin assembly factor-1 required for in vitro interaction with chicken histone deacetylase-2"J. Biol. Chem.. 274. 16646-16653 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Takami, Y., Kikuchi, H. and Nakayama, T.: "Chicken histone deacetylase-2 controls the amount of the IgM H-chain at the steps of both transcription of its gene and alternative processing of its pre-mRNA in the DT40 cell line"J. Biol. Chem.. 274. 23977-23990 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Takechi, S. and Nakayama, T.: "Sas3 is a histone acetyltransferase and requires a zinc finger motif"Biochem. Biophys. Res. Commun.. 266. 405-410 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Nakayama, T., Takami, Y. and Ahmad, A.: "Interaction of the p48 subunit of chicken chromatin assembly factor-1 with histone deacetylases"Current Topics in Biochemical Research. 1. 173-178 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Takami, Y., Nishi, R. and Nakayama, T.: "Histone H1 variants play individual roles in transcription regulation in the DT40 chicken B cell line"Biochem. Biophys. Res. Commun.. (in press). (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Takami, Y. and Nakayama, T.: "N-terminal region, C-terminal region, nuclear export signal and deacetylation activity of histone deacetylase-3 are essential for the viability of the DT40 chicken B cell line"J. Biol. Chem.. (in press). (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Nakayama, T. and Takami, Y.: "Histone deacetylase-2 controls the amount of the IgM H-chain at the steps of both transcription and alternative pre-mRNA processing in the DT40 cell line"Current Topics in Biochemical Research. (in press). (2000)

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

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Published: 2001-10-23  

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