1994 Fiscal Year Annual Research Report

真核生物組換え蛋白質の機能解析

Research Project

Project/Area Number	06044142
Research Institution	Osaka University
Principal Investigator	小川智子大阪大学, 理学部, 講師 (80028208)
Co-Investigator(Kenkyū-buntansha)	コワルコスキーステファカリフォルニア大学, 微生物学部門, 教授エーゲルマンエドワードミネソタ大学, 細胞生物学部門, 助教授ピショップダグラスシカゴ大学, 細胞放射線・腫瘍部門, 講師篠原彰大阪大学, 理学部, 助手 (00252578) 小川英行大阪大学, 理学部, 教授 (70028207)
Keywords	減数分裂期組換え / Rad51蛋白質 / Lim15(Dmc1)蛋白質 / 組換え開始機構 / 染色体構造 / DNA二重鎖切断 / 酵母組換え遺伝子 / ユリ組換え蛋白質
Research Abstract	遺伝的組換えは真核生物で、減数分裂期組換え、抗体遺伝子の再編成、細胞周期の制御に関わり、生物の多様性、遺伝子発現制御に重要な役割を担っている。平成6年度の共同研究では、減数分裂期組換えに直接関与し、しかも、構造が大腸菌RecAと類似のRad51とDmc1蛋白と組換え開始機構について主に解析した。 1 組換え遺伝子産物の発現時期と減数分裂期の染色体上での分布を免疫学的手法で解析し、その動的挙動をユリ、酵母、マウスで解析した。特に染色体構造で減数分裂期の進行が明確にできるユリを用いて、Rad51、Lim15(Dmc1 homolog)がレプトテン期で同じ位置に存在し、パキテン期ではRad51が異なった様式で染色体上にあることを示した(小川智子)。 2 酵母ではRad51がDmc1の染色体への結合に必要であることを示した(Bishop)。 3 Rad51とDmc1蛋白の精製、生化学的活性の検出を行った。また、これらの二重変異株のDNA鎖切断の修復と組換え、染色体構造の変化の関係を解析し、減数分裂期組換えでこれら類似蛋白質の役割分担の解明に迫った(篠原、Bishop)。 4 組換え開始にMRE11、RAD50、XR2遺伝子が複合体を形成して二重鎖切断を行うこと、RNA結合蛋白であるMre2とMerlがMer2RNAのSplicingを行って、減数分裂期組換えの開始を制御していること等を明らかにした(小川英行)。 5 組換え中間体のホリデイ構造の解離に関わるRuvB蛋白とDNAが結合した電子顕微鏡写真のイメージでRuvB蛋白の立体構造を解析した(Egelman)。 6 大腸菌での組換え開始の研究を精力的に行っているKowalzckowski博士(米国)が特別研究協力者として来日し、組換え開始機構の一般性について討議した。

Research Products

(24 results)

All Other

All Publications (24 results)

[Publications] Hitoshi Kurumizaka,Shukuko Ikawa,Tomoatsu Ikeya,Tomoko Ogawa and Takehiko Shibata: "A chimera RecA protein exhibits altered double-stranded DNA-binding." J.Biol.Chem.269. 3068-3075 (1994)
[Publications] Tomoko Ogawa,Akira Shinohara,Akira Nabetani,Tomoatsu Ikeya,Xiong Yu.Edward H.Egelman and Hideyuki Ogawa: "RecA-like Recombination Proteins in Eukaryotes:Functions and Structures of RAD51 Genes." Cold Spring Harbor Symp.Quant.Biol.58. 567-576 (1994)
[Publications] Hitoshi Kurumizaka,B.J.Rao,Tomoko Ogawa,Charles m.Radding and Takehiko Shibata,: "A chimeric RecA protein that implicates non-Watson-Crick interaction in homologous pairing" Nucleic Acid Res.22. 3387-3391 (1994)
[Publications] Tomoko Ogawa Akira Shinohara and Tomoatsu Ikeya: "A species-specific interaction of Rad51 and Rad52 proteins" Adv.Biophysics,. 31. 93-100 (1995)
[Publications] Masahiro Terasawa,Akira Shinohara,Yasuo Hotta,Hideyuki Ogawa and Tomoko Ogawa: "Localization of RecA-like Recombination Proteins on Chromosomes of Lily at Various Meiotic Stages." Genes & Development. (in press). (1995)
[Publications] Ryuichi Kato and Hideyuki Ogawa: "An essential gene,ESR1,is required for mitotic cell growth,DNA repair and meiotic recombination in Saccharomyces cerevisiae" Nucleic Acids Research. 22. 3104-3112 (1994)
[Publications] Katsyki Johzuka and Hideyuki Ogawa: "Interaction of Mre11 and Rad5-:Two protein required for DNA repair and meiosis-specific double-strand break formation in Saccharomyces cerevisiae" Genetics. 139(inpress). (1995)
[Publications] Hideyuki Ogawa,Katsuki Johzuka,Takuro Nakagawa,Sunhee,Leem and A.H.Hagiwara: "Functions of the yeast meiotic recombination genes,MRE11 and MRE2" Adv.Biophysics. 31. 67-76 (1995)
[Publications] Kazuhiro Maeshima,Katsumi Morimatsu,Akira Shinohara and Toshihiro Horii: "Rad51 homologues in Xenopus laevis:two distinct genes and highly expressed in ovary and testis." Gene. inpress. (1995)
[Publications] Dougras Bishop: "RecA homologs Dmc1 and Rad51 interact to form multiple nuclear complexes before meiotic chromosome synapsis." Cell. 79. 1081-1092 (1994)
[Publications] Stasiak,A.and Egelman,E.H.: "Structure and Function of RecA-DNA Complexes." Experientia. 50. 192-203 (1994)
[Publications] Bednar,J.,Furrer,P.,Stasiak,A.,Dubochet,J.,Egelman,E.H.and Bates,A.D.: "The twist,writhe and overall shape of supercoiled DNA changeduring counterion induced transition from a loosely to a tightly interwoundsuperhelix.Possible implications for DNA structure in vivo." J.Mol.Biol.235. 825-847 (1994)
[Publications] Orlova,A.,Yu,X.and Egelman.E.H.: "“Three-dimensional Reconstruction of a Co-complex of F-Actin with Antibody Fab Fragments toActin's Amino Terminus." Biophysical Journal. 66. 276-285 (1994)
[Publications] Egelman,E.H.: "The Ghost of Ribbons Past." Current Biology. 4. 79-81 (1994)
[Publications] Stasiak,A.,Tsaneva,I.,West,S.C.Benson,K.,Yu,X and Egelman,E.H.: "The E.coli RuvB Branch Migration Protein Forms Double Hexameric Rings Around DNA" Proc.Natl.Acad.Sci.U.S.A.91. 7618-7622 (1994)
[Publications] Tsuprun,V.,Anderson,D.and Egelman,E.H.: "The Bacteriophage i29 Head-TailConnector Shows 13-Fold Symmetry in Both Hexagonally-Packed Arrays and as Single Particles" Biophysical Journal. 66. 2139-2150 (1994)
[Publications] Egelman,E.H.,Yu,X.,Wild,R.,Hingorani,M.M.and Patel,S.S: "T7 Helicase/Primase Proteins Form Rings Around Single-Stranded DNA that Suggest a General Structure for Hexameric Helicases" Proc.Natl.Acad.Sci.U.S.A. (in press). (1995)
[Publications] Egelman,E.H: "New Insights On F-actin Dynamics." Current Opinion in Structural Biology(invited review). (in press). (1995)
[Publications] Egelann,E.H: "Electron Microscopy of DNA Helicases" Quarterly Review of Biophysics(invited review). (in press). (1955)
[Publications] 小川智子篠原彰小川英行: "真核生物の組換え蛋白質Rad51とRad52の機能と構造" 実験医学(羊土社)「トッピクス」. 12. 524-547 (1994)
[Publications] 小川英行、小川智子: "酵母に観る最新の真核生物像VI.組換え機構減数分裂期組換え-関与する遺伝子とその普遍性-" 蛋白質核酸酵素(共立出版)「臨時増刊号」. 39. 568-578 (1994)
[Publications] 篠原彰小川英行小川智子: "真核生物の組換え(修復)に働く遺伝子" 細胞工学(秀潤社). 13. 710-719 (1994)
[Publications] 小川智子: "真核生物の組換え蛋白質の機能と構造" 第25回放射性総合医学研究シンポジウム. 25. 25-37 (1994)
[Publications] 篠原彰小川英行小川智子: "真核生物RAD51遺伝子の構造と機能" 日本臨床(日本臨床社). 53. 239-249 (1994)

1994 Fiscal Year Annual Research Report

真核生物組換え蛋白質の機能解析

Principal Investigator

小川 智子 大阪大学, 理学部, 講師 (80028208)

Research Products

[Publications] Hitoshi Kurumizaka,Shukuko Ikawa,Tomoatsu Ikeya,Tomoko Ogawa and Takehiko Shibata: "A chimera RecA protein exhibits altered double-stranded DNA-binding." J.Biol.Chem.269. 3068-3075 (1994)

[Publications] Tomoko Ogawa,Akira Shinohara,Akira Nabetani,Tomoatsu Ikeya,Xiong Yu.Edward H.Egelman and Hideyuki Ogawa: "RecA-like Recombination Proteins in Eukaryotes:Functions and Structures of RAD51 Genes." Cold Spring Harbor Symp.Quant.Biol.58. 567-576 (1994)

[Publications] Hitoshi Kurumizaka,B.J.Rao,Tomoko Ogawa,Charles m.Radding and Takehiko Shibata,: "A chimeric RecA protein that implicates non-Watson-Crick interaction in homologous pairing" Nucleic Acid Res.22. 3387-3391 (1994)

[Publications] Tomoko Ogawa Akira Shinohara and Tomoatsu Ikeya: "A species-specific interaction of Rad51 and Rad52 proteins" Adv.Biophysics,. 31. 93-100 (1995)

[Publications] Masahiro Terasawa,Akira Shinohara,Yasuo Hotta,Hideyuki Ogawa and Tomoko Ogawa: "Localization of RecA-like Recombination Proteins on Chromosomes of Lily at Various Meiotic Stages." Genes & Development. (in press). (1995)

[Publications] Ryuichi Kato and Hideyuki Ogawa: "An essential gene,ESR1,is required for mitotic cell growth,DNA repair and meiotic recombination in Saccharomyces cerevisiae" Nucleic Acids Research. 22. 3104-3112 (1994)

[Publications] Katsyki Johzuka and Hideyuki Ogawa: "Interaction of Mre11 and Rad5-:Two protein required for DNA repair and meiosis-specific double-strand break formation in Saccharomyces cerevisiae" Genetics. 139(inpress). (1995)

[Publications] Hideyuki Ogawa,Katsuki Johzuka,Takuro Nakagawa,Sunhee,Leem and A.H.Hagiwara: "Functions of the yeast meiotic recombination genes,MRE11 and MRE2" Adv.Biophysics. 31. 67-76 (1995)

[Publications] Kazuhiro Maeshima,Katsumi Morimatsu,Akira Shinohara and Toshihiro Horii: "Rad51 homologues in Xenopus laevis:two distinct genes and highly expressed in ovary and testis." Gene. inpress. (1995)

[Publications] Dougras Bishop: "RecA homologs Dmc1 and Rad51 interact to form multiple nuclear complexes before meiotic chromosome synapsis." Cell. 79. 1081-1092 (1994)

[Publications] Stasiak,A.and Egelman,E.H.: "Structure and Function of RecA-DNA Complexes." Experientia. 50. 192-203 (1994)

[Publications] Orlova,A.,Yu,X.and Egelman.E.H.: "“Three-dimensional Reconstruction of a Co-complex of F-Actin with Antibody Fab Fragments toActin's Amino Terminus." Biophysical Journal. 66. 276-285 (1994)

[Publications] Egelman,E.H.: "The Ghost of Ribbons Past." Current Biology. 4. 79-81 (1994)

[Publications] Stasiak,A.,Tsaneva,I.,West,S.C.Benson,K.,Yu,X and Egelman,E.H.: "The E.coli RuvB Branch Migration Protein Forms Double Hexameric Rings Around DNA" Proc.Natl.Acad.Sci.U.S.A.91. 7618-7622 (1994)

[Publications] Tsuprun,V.,Anderson,D.and Egelman,E.H.: "The Bacteriophage i29 Head-TailConnector Shows 13-Fold Symmetry in Both Hexagonally-Packed Arrays and as Single Particles" Biophysical Journal. 66. 2139-2150 (1994)

[Publications] Egelman,E.H.,Yu,X.,Wild,R.,Hingorani,M.M.and Patel,S.S: "T7 Helicase/Primase Proteins Form Rings Around Single-Stranded DNA that Suggest a General Structure for Hexameric Helicases" Proc.Natl.Acad.Sci.U.S.A. (in press). (1995)

[Publications] Egelman,E.H: "New Insights On F-actin Dynamics." Current Opinion in Structural Biology(invited review). (in press). (1995)

[Publications] Egelann,E.H: "Electron Microscopy of DNA Helicases" Quarterly Review of Biophysics(invited review). (in press). (1955)

[Publications] 小川智子 篠原 彰 小川英行: "真核生物の組換え蛋白質Rad51とRad52の機能と構造" 実験医学(羊土社)「トッピクス」. 12. 524-547 (1994)

[Publications] 小川英行、小川智子: "酵母に観る最新の真核生物像VI.組換え機構 減数分裂期組換え-関与する遺伝子とその普遍性-" 蛋白質 核酸 酵素(共立出版)「臨時増刊号」. 39. 568-578 (1994)

[Publications] 篠原 彰 小川英行 小川智子: "真核生物の組換え(修復)に働く遺伝子" 細胞工学(秀潤社). 13. 710-719 (1994)

[Publications] 小川智子: "真核生物の組換え蛋白質の機能と構造" 第25回 放射性総合医学研究シンポジウム. 25. 25-37 (1994)

[Publications] 篠原 彰 小川英行 小川智子: "真核生物RAD51遺伝子の構造と機能" 日本臨床(日本臨床社). 53. 239-249 (1994)

小川智子大阪大学, 理学部, 講師 (80028208)

[Publications] 小川智子篠原彰小川英行: "真核生物の組換え蛋白質Rad51とRad52の機能と構造" 実験医学(羊土社)「トッピクス」. 12. 524-547 (1994)

[Publications] 小川英行、小川智子: "酵母に観る最新の真核生物像VI.組換え機構減数分裂期組換え-関与する遺伝子とその普遍性-" 蛋白質核酸酵素(共立出版)「臨時増刊号」. 39. 568-578 (1994)

[Publications] 篠原彰小川英行小川智子: "真核生物の組換え(修復)に働く遺伝子" 細胞工学(秀潤社). 13. 710-719 (1994)

[Publications] 小川智子: "真核生物の組換え蛋白質の機能と構造" 第25回放射性総合医学研究シンポジウム. 25. 25-37 (1994)

[Publications] 篠原彰小川英行小川智子: "真核生物RAD51遺伝子の構造と機能" 日本臨床(日本臨床社). 53. 239-249 (1994)