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1998 Fiscal Year Final Research Report Summary

Structural and Functional Analyzes of Human Artificial Chromosomes

Research Project

Project/Area Number 09480185
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Molecular biology
Research InstitutionNagoya University

Principal Investigator

MASUMOTO Hiroshi  Nagoya University, Graduate School of Science, Assistant Professor, 大学院・理学研究科, 講師 (70229384)

Co-Investigator(Kenkyū-buntansha) NOZAKI Naohito  Kanagawa Dental College, Research Associate, 歯学部, 助手 (70222198)
Project Period (FY) 1997 – 1998
Keywordscetromere / teloreme / alphoid DNA / YAC / HAC / CENP-B
Research Abstract

In order to define a functional human centromere sequence, an artificial chromosome was constructed as a reproducible DNA molecule. Mammalian telomere repeats and a selectable marker were introduced into yeast artificial chromosomes (YACs) containing alphoid DNA from the human chromosome 21 centromere region in a recombination-deficient yeast host. When these modified YACs were introduced into human cultured cells, a YAC with the alphoid DNA from alpha21-I locus, containing CENP-B boxes at a high frequency and a regular repeat array, efficiently formed artificial chromosomes which were maintained stably in the absence of selection and bound centromere/kinetochore proteins, CENP-A, CENP-B, CENP-C and CENP-E.The artificial chromosomes 1 - 5 Mb in size and composed of multimers of the introduced YAC DNA, aligned at metaphase plates and segregated to opposite poles correctly in anaphase. Extensive cytological analyses strongly suggested that they had not acquired host sequences and were formed in all cases by a de novo mechanism. In contrast, artificial chromosomes were never produced with a modified YAC containing alphoid DNA from the alpha21-II locus which contains no CENP-B boxes and has a less regular sequence arrangement. We conclude that alpha21-I alphoid DNA can induce de novo assembly of active centromere/kinetochore structures on artificial chromosomes.

  • Research Products

    (6 results)

All Other

All Publications (6 results)

  • [Publications] J.Iwahara: "A helix-turn-helix structure unit in human centromere protein B (CENP-B)." EMBO J.17. 827-837 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] M.Ikeno: "Construction of YAC-based mammalian artificial chromosomes." Nature Biotech.16. 431-439 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] H.Masumoto: "Assay of centromere function using a human artificial chromosome." Chromosoma. 107. 406-416 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] J.Iwahara et al.: "A helix-turn-helix structure unit in human centromere protein B (CENP-B)." EMBO J.17. 827-837 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] M.Ikeno et al.: "Construction of YAC-based mammalian artificial chromosomes." Nature Biotech.16. 431-439 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] H.Masumoto et al.: "Assay of centromete function using a human artificial chromosome." Chromosoma. 107. 406-416 (1998)

    • 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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