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2003 Fiscal Year Final Research Report Summary

Investigation on functions of genes on the Z and W sex chromosomes of birds with respect to sex determination and sex differentiation

Research Project

Project/Area Number 12460036
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field 応用微生物学・応用生物化学
Research InstitutionNihon University

Principal Investigator

MIZUNO Shigeki  Nihon University, College of Bioresource Sciences, Department of Agricultural and Biological Chemistry, Professor, 生物資源科学部, 教授 (90112903)

Co-Investigator(Kenkyū-buntansha) OGIHARA Jun  Nihon University, College of Bioresource Sciences, Department of Agricultural and Biological Chemistry, Lecturer, 生物資源科学部, 講師 (50256830)
KATO Jun  Nihon University, College of Bioresource Sciences, Department of Agricultural and Biological Chemistry, Lecturer, 生物資源科学部, 講師 (80130467)
Project Period (FY) 2000 – 2003
Keywordschicken / W chromosome / Z chromosome / genes on sex chromosomes / sex determination / polypurine / polypyrimidine sequence / non-coding RNA / DMRT1 gene
Research Abstract

(1) cDNA clones of a novel W chromosome-linked gene, PKCI-W, and its counterpart gene on the Z chromosome, PKCI-Z, were obtained from the female-minus-male subtraction cDNA library from day-S chicken embryos. Homology of PKCI-W and PKCI-Z at the amino acid sequence level was about 60%. The mRNA-level expression of both genes was high in genital ridge and neural tissues in early female embryos. PKCI-Z was a chicken ortholog of mammalian PKCI/HINT. PKCI-W lacked a HIT (histidine triad) motif which is essential for the function of PKCI/HINT but regions flanking the HIT motif which are involved in the homodimer formation of PKCI/HINT were well conserved. Both PKCI-W and PKCI-Z were expressed as fusion proteins with glutathione S-transferase or maltose-binding protein in E. coli and purified. PKCI-Z showed high affinity binding to adenosine and AMP-amidate hydrolyzing activity as demonstrated for mammalian PKCI/HINT, whereas PKCI-W did not show these activities. These activities of PKCI-Z w … More ere inhibited in a dominant-negative manner by forming a heterodimer with PKCI-W in vitro. These results supported the model in which PKCI-W might participate in the female sex determination.
(2) Genomic clones for spindlin gene (SPIN-W) and a novel SspI-family repetitive sequence were obtained from the chicken W chromosome-specific genomic library which was prepared by laser microdissection of a metaphase chromosome set followed by random PCR amplification. A clone for the counterpart gene, SPIN-Z, on the Z chromosome was subsequently obtained. Northern blot analysis using the 3'-untranslated region probe revealed that SPIN-Z was expressed ubiquitously but SPIN-W was expressed predominantly in the ovary. The SspI-family consisted of 11,300 tandem repeats of a 508-bp unit and localized at the euchromatin/ heterochromatin boundary on the short arm of the chicken W chromosome. Each repeating unit contained a 120-bp polypurine/polypyrimidine sequenece which might form a triple-stranded DNA. The SspI-family formed a diffuse chromatin structure in nuclei of female chicken embryonic fibroblasts.
(3) A clone for a specific region on the Z chromosome, designated MIIM (male-hypermethylated) region, was obtained from the chicken oocyte cDNA library. In females, MIL-IM region was transcribed actively into about 9.5-kb non-coding RNA (MHM-RNA) which accumulated at or in the close vicinity of the MHM region in a nucleus. In males, MHM region was highly methylated and its transcription was totally inactive. MHM-RNA accumulated very close to the DMRTI gene in a nucleus. The DMRT1 gene, which has been suggested to be essential for testis differentiation, is not expressed in female chickens after hatching and MHM-RNA was speculated to be involved in this transcriptional inactivation. Less

  • Research Products

    (18 results)

All Other

All Publications (18 results)

  • [Publications] S.Mizuno: "Z and W chromosomes of chickens : studies on their gene functions in sex determination and sex differentiation"Cytogenetics and Genome Research. 99. 236-244 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] J.Kato: "Efficient selection of genomic clones from a female chicken bacterial artificial chromosome library by four-dimensional polymerase chain reactions"Poultry Sciences. 81. 1501-1508 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Y.Itoh: "Molecular and cytological characterization of SspI-family repetitive sequence on the chicken W chromosome"Chromosome Research. 10. 499-511 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Y.Kuroda: "Absence of Z-chromosome inactivation for five genes in male chickens"Chromosome Research. 9. 457-468 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Y.Itoh: "Chicken spindlin genes on W and Z chromosomes : transcriptional expression of both genes and dynamic behavior of spindlin in interphase and mitotic cells"Chromosome Research. 9. 283-299 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] T.Hori: "Wpkci, encoding an altered form of PKCI, is conserved widely on the avian W chromosome and expressed in early female embryos : implication of its role in female sex determination"Molecular Biology of the Cell. 11. 3645-3660 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] M.Teranishi, S.Mizuno: "The male hypermethylation (MHM) region on the chicken Z chromosome : female-specific transcription and its biological implication."Chromosomes Today. 14. 45-53 (2004)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] S.Mizuno, R.Kunita, O.Nakabayashi, Y.Kuroda, N.Arai, M.Harata, A.Ogawa, Y.Itoh, M.Teranishi, T.Hori: "Z and W chromosomes of chickens : studies on their gene functions in sex determination and sex differentiation."Cytogenetics and Genome Research. 99. 236-244 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] J.Kato, T.Hattori, S.Ohba, Y.Tamaki, N.Yamada, T.Taguchi, J.Ogihara, K.Ohya, Y.Itoh, T.Hori, S.Asakawa, N.Shimizu, S.Mizuno: "Efficient selection of genomic clones from a female chicken bacterial artificial chromosome library by four-dimensional polymerase chain reactions."Poultry Sciences. 81. 1501-1508 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Y.Itoh, S._Mizuno: "Molecular and cytological characterization of SspI-family repetitive sequence on the chicken W chromosome."Chromosome Research. 10. 499-511 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Y.Itoh, M.Suzuki, A.Ogawa, I.Munechika, K.Murata, S.Mizuno.: "Identification of the sex of a wide range of Carinatae birds by PCR using primer sets selected from chicken EEO.6 and its related sequences."Journal of Heredity. 92. 315-321 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Y.Kuroda, N.Arai, M.Arita, M.Teranishi, T.Hori, M.Harata, S.Mizuno: "Absence of Z-chromosome inactivation for five genes in male chickens."Chromosome Research. 9. 457-468 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Y.Itoh, T.Hori, H.Saitoh, S.Mizuno: "Chicken spindlin genes on W and Z chromosomes : transcriptional expression of both genes and dynamic behavior of spindlin in interphase and mitotic cells."Chromosome Research. 9. 283-299 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] M.Teranishi, Y.Shimada, T.Hori, O.Nakabayashi, T.Kikuchi, T.Maclead, R.Pym, B.Sheldon, I.Solovei, H.Macgregor, S.Mizuno: "Transcripts of the MHM region on the chicken Z chromosome accumulate as non-coding RNA in the nucleus of femele cells adjacent to the DMRT1 locus."Chromosome Research. 9. 147-165 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] S.Mizuno: "Current studies on mechanisms of sex determination and sex differentiation with the ZW sex chromosomes of birds: comparison with studies on mammalian systems (in Japanese)."Seikagaku. 73. 1411-1427 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] I.Nanda, B.Zend-Ajusch, Z.Shan, F.GrUtzner, M.Scharti, D.W.Burt, M.Koehler, V.M.Fowler, G.Goodwin, W.J.Schneider, S.Mizuno, O.Dechant, T.Haaf, M.Sthmid: "Conserved synteny between the chicken Z sex chromosome and human chromosome 9 includes the male regulatory gene DMRT1: a comparative (re)view on avian sex determination."Cytogenetics and Cell Genetics. 89. 67-78 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] T.Hori, S.Asakawa, Y.Itoh, N.Shimizu, S.Mizuno.: "Wpkci, encoding an altered form of PKCI, is conserved widely on the avian W chromosome and expressed in early female embryos : implication of its role in female sex determination."Molecular Biology of the Cell. 11. 3645-3660 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] M.Schmid, I.Nanda, M.Guttenbach, C.Steinlein, H.Hoehn, M.Schartl, T.Haaf, S.Weigend, R.Fries, J-M.Buerstedde, K.Wimmers, D.W.Burt, J.Smith, S.A'Hara, A.Law, D.K.Griffin, N.Bumstead, J.Kaufman, A.Thomson, T.Burke, M.A.M.Groenen, R.M.A.Crooijmans, A.Vignal, V.Fillon, M.Morisson, F.Pitel, M.Tixier-Boichard, K.Ladjali-Mohammedi, J.Hillel, A.Maki-Tanila, H.H.Cheng, M.E.Delany, J.Burnside, S._Mizuno: "First report on chicken genes and chromosomes 2000."Cytogenetics and Cell Genetics. 90. 169-218 (2000)

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

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Published: 2005-04-19  

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