1992 Fiscal Year Annual Research Report

組換え機構の分子的基礎

Research Project

Project/Area Number	04262102
Research Institution	Nagoya University
Principal Investigator	堀田康雄名古屋大学, 理学部, 教授 (30190218)
Co-Investigator(Kenkyū-buntansha)	城石俊彦国立遺伝学研究所, 細胞遺伝研究系, 助手 (90171058) 柴田武彦理化学研究所, バイオデザイン研究グループ, 主任研究員 (70087550) 小林一三東京大学, 医科学研究所, 助教授 (30126057) 大坪栄一東京大学, 応用微生物研究所, 教授 (10158800) 小川英行大阪大学, 理学部, 教授 (70028207)
Keywords	遺伝子組換え / 相同組換え / 非相同組換え / 減数分裂 / ホットスポット / RAD51 / RecA / 核骨格
Research Abstract	酵母の遺伝子組換えに働くRAD51ホモログを、ヒト・マウス・鶏で同定し、遺伝子のクロ-ン化・蛋白の一次構造を明らかにした。RAD51はヒトでは15番染色体上に、マウスでは2番染色体にあり、脾臓・胸腺・卵巣で高い発現を示す(小川英行)。RAD51の機能解析よりRecA蛋白と高い類似性(反応要求性・ATPase・RAD51-ATP-dsDNA複合体の活性)を認定し、組換え蛋白の普遍性を示唆した(小川智子)。大腸菌と緑膿菌のキメラRecA蛋白の解析から、RecA蛋白のDNA結合領域の特異性を示した(柴田)。相同組換え領域端で第3のDNA分子との非相同組換えを示す産物をマウス細胞と大腸菌で同定し、相同組換えが非相同組換えを誘起することを示した(小林)。減数分裂の遺伝子組換えにはホットスポットと組換え促進因子と抑制因子が関与する。野生マウス由来wm7染色体上の抑制因子はMHC領域内ホットスポットとテロメア側Cyp21遺伝子間に座し第17染色体の20メガベ-ス以上の領域の組換えを、雄に於て抑制する。促進因子に対しCisでもtransでも抑制を起す(城石)。ユリ減数分裂特異的発現遺伝子(LIM1〜LIM18)の構造解析を完了し、その1つが酵母のDMC1であり、さらに別の相似遺伝子を同定し、共に減数分裂に不可欠であることを示した(堀田)。マウス細胞で4種のDNA鎖転移活性を見いだしCHO細胞の欠失aprt遺伝子を標的として、組換えが1本鎖でも2本鎖でも同効率で起こることを示した(小山)。IS3はIS1と同じくフレ-ムシフトでトランスポ-ゼスを生産し、一塩基挿入後トランスポ-ゼスを生産させると多量の欠失変異体と転移中間体分子を生産する(大坪)。減数分裂特異的調節因子と核骨格蛋白が染色体対合に貢献することを示唆し(堀田)、減数分裂時組換え制御の分子レベルでの機構解析を伸展させ、これらの生物界での普遍性と体細胞での組換えとの差異を明らかにした。さらに多くの研究すベき問題が出てきた。

Research Products
(19 results)

All Other

All Publications (19 results)

[Publications] Furukawa,K.: "cDNA cloning of a germ cell specific laminB3 from mouse spermatocyte and functional analysis by ectopic expression in somatic cultured cells." EMBO J.12. 97-106 (1993)
[Publications] Ooyama,T.: "Isolation of synaptonemal conplexes from lily microsporocytes." Plant Science. 86. 115-124 (1992)
[Publications] Kobayashi,T.: "Isolation and characterization of yeast gene which is homologous with meiosis-specific cDNA from a plant." Molec.Gen.genet.(1992)
[Publications] Shinohara,A.: "Rad51 Protein Involved in Repair and Recombi nation in S.cerevisiae Is aRecA-like Protein." Cell. 69. 457-470 (1992)
[Publications] Leem S H.: "The MRE4 gene encodes a novel protein kinase homologue required for meiotic recombination in Saccharomyces cerevisiae." Nucleic Acids Research. 20. 449-457 (1992)
[Publications] Sekine,Y.: "Identification of the site of translational frameshifting required for production of the transposase encoded by insertion sequence IS1." Mol.Gen.Genet.235. 317-324 (1992)
[Publications] Sekine,Y.: "DNA sequences requied for translational frameshifting in produciton of the trans-posase encoded by IS1." Mol.Gen.Genet.235. 325-332 (1992)
[Publications] Mishina,Y.: "Identification of two types of homologous DNA pairing activity in mouse cells." Biochem.Biophys.Res.Commun.185. 987-992 (1992)
[Publications] Aratani,Y.: "End extension repair of introduced targeting vectors mediated by homologous recomgination in mammalion cells." Nucleic Acid Res.20. 4795-4801 (1992)
[Publications] Kobayashi,I.: "Mechanisms for gene conversion and homologous recombination." Advances in Biophysics. 28. 81-133 (1992)
[Publications] Takahashi,N.: "Nonconservative recombination in Escherichia coli." Proc.Natl.Acad.Sci.89. 5912-5916 (1992)
[Publications] Yamamoto,K.: "Gene conversion in the Escherichia coli RecF pathway." Mol.Gen.Genet.234. 1-13 (1992)
[Publications] Ikeda,M.: "Epitope mapping of anti-recA protein IgGs by region specified polymerase chain reaction mutagenesis." J.Biol.Chem.267. 6291-6296 (1992)
[Publications] Arai,N.: "Phenol-treatment and a homologous pairing-assay." J.Biol.Chem.267. 3679-3684 (1992)
[Publications] Mishina,Y.: "Identification of two types of homologous DNA pairing sctivity in mouse cells." Biochem.Biophys.Res.Commum.185. 987-992 (1992)
[Publications] Ogawa,T.: "Functional Structure of the RecA Protein Found Chimera Analysis." J.Mol.Biol.226. 651-660 (1992)
[Publications] Ogawa,T.: "Similarity of the Yeast Rad51 Filament to the Bacterial RecA Filament." Science. (1993)
[Publications] Koide,T.: "The amplified long genomic sequence(ALGS)located in the centromeric regions of mouse chromosomes." Genomics. 13. 1186-1191 (1992)
[Publications] Shiroishi,T.: "Hotspots of homologous recombination in the mouse major histocompatibility complex." Genetica. (1993)

1992 Fiscal Year Annual Research Report

組換え機構の分子的基礎

Principal Investigator

堀田 康雄 名古屋大学, 理学部, 教授 (30190218)

Research Products

[Publications] Furukawa,K.: "cDNA cloning of a germ cell specific laminB3 from mouse spermatocyte and functional analysis by ectopic expression in somatic cultured cells." EMBO J.12. 97-106 (1993)

[Publications] Ooyama,T.: "Isolation of synaptonemal conplexes from lily microsporocytes." Plant Science. 86. 115-124 (1992)

[Publications] Kobayashi,T.: "Isolation and characterization of yeast gene which is homologous with meiosis-specific cDNA from a plant." Molec.Gen.genet.(1992)

[Publications] Shinohara,A.: "Rad51 Protein Involved in Repair and Recombi nation in S.cerevisiae Is aRecA-like Protein." Cell. 69. 457-470 (1992)

[Publications] Leem S H.: "The MRE4 gene encodes a novel protein kinase homologue required for meiotic recombination in Saccharomyces cerevisiae." Nucleic Acids Research. 20. 449-457 (1992)

[Publications] Sekine,Y.: "Identification of the site of translational frameshifting required for production of the transposase encoded by insertion sequence IS1." Mol.Gen.Genet.235. 317-324 (1992)

[Publications] Sekine,Y.: "DNA sequences requied for translational frameshifting in produciton of the trans-posase encoded by IS1." Mol.Gen.Genet.235. 325-332 (1992)

[Publications] Mishina,Y.: "Identification of two types of homologous DNA pairing activity in mouse cells." Biochem.Biophys.Res.Commun.185. 987-992 (1992)

[Publications] Aratani,Y.: "End extension repair of introduced targeting vectors mediated by homologous recomgination in mammalion cells." Nucleic Acid Res.20. 4795-4801 (1992)

[Publications] Kobayashi,I.: "Mechanisms for gene conversion and homologous recombination." Advances in Biophysics. 28. 81-133 (1992)

[Publications] Takahashi,N.: "Nonconservative recombination in Escherichia coli." Proc.Natl.Acad.Sci.89. 5912-5916 (1992)

[Publications] Yamamoto,K.: "Gene conversion in the Escherichia coli RecF pathway." Mol.Gen.Genet.234. 1-13 (1992)

[Publications] Ikeda,M.: "Epitope mapping of anti-recA protein IgGs by region specified polymerase chain reaction mutagenesis." J.Biol.Chem.267. 6291-6296 (1992)

[Publications] Arai,N.: "Phenol-treatment and a homologous pairing-assay." J.Biol.Chem.267. 3679-3684 (1992)

[Publications] Mishina,Y.: "Identification of two types of homologous DNA pairing sctivity in mouse cells." Biochem.Biophys.Res.Commum.185. 987-992 (1992)

[Publications] Ogawa,T.: "Functional Structure of the RecA Protein Found Chimera Analysis." J.Mol.Biol.226. 651-660 (1992)

[Publications] Ogawa,T.: "Similarity of the Yeast Rad51 Filament to the Bacterial RecA Filament." Science. (1993)

[Publications] Koide,T.: "The amplified long genomic sequence(ALGS)located in the centromeric regions of mouse chromosomes." Genomics. 13. 1186-1191 (1992)

[Publications] Shiroishi,T.: "Hotspots of homologous recombination in the mouse major histocompatibility complex." Genetica. (1993)

堀田康雄名古屋大学, 理学部, 教授 (30190218)