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1997 Fiscal Year Final Research Report Summary

Methods to analyze nuclear organizition of mammalian chromosomes in interphase nuclei

Research Project

Project/Area Number 08554033
Research Category

Grant-in-Aid for Scientific Research (A)

Allocation TypeSingle-year Grants
Section展開研究
Research Field 遺伝
Research InstitutionNATIONAL INSTITUTE OF GENETICS

Principal Investigator

IKEMURA Toshimichi  National Inst.of Genetics, Dept.of Popul.Biol., Professor, 集団遺伝研究系, 教授 (50025475)

Co-Investigator(Kenkyū-buntansha) KAMIYA Kiyoshi  Hamamatsu Photonics, Head of Sistems Division., システム事業部, 部長
OKUMURA Katsuzumi  Mie University, Faculty of Bioresources, Assistant Professor, 生物資源学部, 助教授 (30177183)
Project Period (FY) 1996 – 1997
Keywordsnon-B DNA structure / centromere / telomere / triple-helix / chromosome / trinucleotide expansion / nuclear organization / multi-color FISH
Research Abstract

Ordered arrangement of chromosomal DNAs into specific domains in nuclei has been proposed to fulfilll important roles in proper gene expression and DNA replication, as well as recombination especially in meiotic nuclei. Formation of non-B DNA structures such as triplex is thought important as a molecular mechanism determining spatial organization in nuclei. Triplex formation occurs readily in polypouine/polypyrimidine sequences leaving single-strand unpaired DNAs that can hybridize with other single-strand DNAs with significant complementarity. This will enable distantly spaced DNAs in genome sequences to associate with each other in interphase nuclei. Distantly spaced DNAs may also form transmolecular triplexes organizing themselves into an ordered array. To investigate mechanisms governing spatial organization of chromosomal DNAs in human and mouse nuclei, we develop multi-colors in situ binding assay methods for DNA probes with specific characteristics such as triplex formation, both in human mitotic and mouse meiotic nuclei. The method resembled fluorescence in situ hybridization used for detecting single-strand RNAs, thus was able to detect single-strand DNAs in nondenatured nuclei. Polyporine/polypyrimidine sequences, such as (GA/TC)n, (GAA/TTC)n and those present in human and mouse genomes, gave clear binding signals with different probe sequences resulting in different signal patterns. Using five pairs of filters optimized for excitation and emission lights that were controlled by a microcomputer, 5 colors of fluorescences were taken without mixing the colors with a cooled CCD camera.

  • Research Products

    (18 results)

All Other

All Publications (18 results)

  • [Publications] 深川 滝郎: "Human pseudoautosomal boundary-like sequences(PABLs):Core and consensus sequence, expression, and involvement in formation of the present day pseudoautosomal boundary of short arm of human sex chromosomes." Human Molecular Genetics. 5. 23-32 (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 安藤 麻子: "cDNA cloning of the human homologues of the mouse Ke4 and Ke6 genes at thc centromeric end of the human MHC region." Geno,mics. 35. 600-602 (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 笠原 正典: "Chromosomal localization of the proteasome Z subunit gene reveals an ancient chromosomal duplication involving the major histocompatibiliy complex." Proc.Natl.Acad.Sci(USA). 93. 9096-9101 (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 菅谷 公彦: "Gene organization of human NOTCH4 and(CTG)n polymorphism in this human counterpart gene of mouse proto-oncogene Int3." Gene. 189. 235-244 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 天前 豊明: "Precisc switching of DNA replication timing in the GC content transition area in the human major histocompatibility complex." Mol.Cell.Biol.17. 4043-4050 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 安藤 麻子: "Triplet repeat polymorphism in the NOTCH4 gene within the human major histocompatibility complex in a healthy population and patients with a salivary gland tumor in Japan." Tissue Antigens. 50. 66-70 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 安藤 麻子: "Triplet repeat polymorphism with in the NOTCH4 gene located near the junction of the HLA class II and class III regions in narcolepsy." Tissue Antigens. 50. 646-649 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 椎名 隆: "Nuclcetide sequencing analysis of the 146kb segment around the IKBL and MICA genes at the centromeric end of the HLA class 1 region." Geno'mics. (in press). (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 大野 みずき: "Affinity sites for triplex DNAs in human nuclei." (投稿中).

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Tatsuo Fukagawa: "Human pseudoautosomal boundary-like sequences (PABLs) : Core and consensus sequence, expression, and involvement in formation of the present day pseudoautosomal boundaty of short arm of human sex chromosomes." Hum.Mol.Genet.5. 23-32 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Asako Ando: "cDNA cloning of the human homologues of the mouse Ke4 and Ke6 genes at the centromeric end of the human MHC region." Genemics. 35. 600-602 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Masanori Kasahara: "Chromosomal localization of the proteasome Z subunit gene reveals an ancient chromosomal duplication involving the major histocompatibiliy complex." Proc.Natl.Acad.Sci.(USA). 93. 9096-9101 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kimihiko Sugaya: "Gene organization of human NOTCH4 and (CTG)n polymorphism im this human conuterpart gene of mouse proto-oncogene Int3." Gene.189. 235-244 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Toyoaki Tenzen: "Precise switching of DNA replication timing in the GC content transition area in the human major histocompatibility complex." Mol.Cell.Biol.17. 4043-4050 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Asako Ando: "Triplet repeat polymorphism in the NOTCH4 gene within the human major histocompatibility complex in a healthy population and patients with a salivary gland tumor in Japan." Tissue Antigens. 50. 66-70

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Asako Ando: "Triplet repeat polymorphism within the NOTCH4 gene located near the junctio of the HLA class II and class III regions in narcolepsy." Tissue Antigens. 50. 646-649

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Takashi Shiina: "Nucleotide sequencing analysis of the 146 Kb segment around the IKBL and MICA genes at the centromeric end of the HLA class 1 region." Genomics. (in press.).

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Mizuki Ohno: "Affinity sites for triplex DNAs in human nuclei." (under description).

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

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Published: 1999-03-16  

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