1993 Fiscal Year Annual Research Report

ゲノムの複製終結と分配制御

Research Project

Project/Area Number	04256103
Research Institution	Kumamoto University
Principal Investigator	平賀壮太熊本大学, 医学部, 教授 (40027321)
Co-Investigator(Kenkyū-buntansha)	小椋光熊本大学, 医学部, 助教授 (00158825) 西村昭子国立遺伝学研究所, 助教授 (20142002) 加藤潤一東京大学, 医科学研究所, 助手 (10194820) 丹羽修身京都大学, 理学部, 助教授 (70144318) 岡崎恒子名古屋大学, 理学部, 教授 (10022584)
Keywords	MukB蛋白 / トポイソメラーゼ / FtsH蛋白 / ヒト染色体 / セントロメア / アルフォイドDNA / 分裂酵母 / ミニ染色体
Research Abstract	1.大腸菌染色体分配に関与するMukB蛋白を精製して、この蛋白の2量体はN端とC端に球状ドメインを持ち中心部がコイルドコイルのロッド状をしていることを電顕で明かにした。またMuKB蛋白はDNA結合能、ATP/GTP結合能を持つことを証明した。mukBオペロンの構成を調べ、mukB遺伝子の上流に細胞の致死に関与する二つの遺伝子(kicB,kicAと命名)が存在することを明らかにした。またmukB106変異のマルチコピーサプレッサーを解析して、コールドショック蛋白とホモロジーを持つ二つの遺伝子を同定した(平賀)。 4種のトポイソメラーゼの二、三、四重変異株を解析して、in vivoでは完全に複製が終わったプラスミドのカテナンの分離はtopoIVによって行われるが、複製が完了する前のプラスミドの分離の経路もあり、そこではtopoII(gyrase)やtopoIIIも働きうることが示唆された(加藤)。 3.単離したマウスのtopoIIalphaのDNAクローンは、酵母のtopoIIの欠失変異を相補することができるが、この系を使いマウスtopoIIalphaのts変異株を分離し機能ドメインの解析を行った(菊地)。 4.染色体分配異常par-271変異はgyrG(CTP合成酵素の遺伝子)の変異であることを明らかにした(西村(行))。 5.FtsH蛋白を精製し、約90%の純度の標品を得た。この標品はATP結合能、ATPase活性を示した(小椋)。 6.ヒストン様蛋白H-NSの変異株では高頻度で無核細胞が放出されるが、この原因は染色体DNAの複製開始頻度が低下したにもかかわらず、細胞分裂との共役性が失われたためであるということが示唆された。またfcsA29変異株では高濃度のカルシウムによって細胞分裂が阻害され巨大細胞を形成して死に至ることを見いだした(永井)。 7.ヒト染色体セントロメア領域に存在するアルフォイドDNA中の17bp配列(CEN-B-Box)にCENP-B蛋白が特異的に結合することを明かにし、この領域のDNA-蛋白複合体の高次構造についてのモデルを提出した(岡崎)。 8.分裂酵母の環状ミニ染色体の脱落頻度を増加させる変異sta1遺伝子を単離し、紡錘体の形成に関わるsadI遺伝子と同一であることを明らかにした(丹羽)。 9.過剰な細胞分裂によって有核のミニ細胞となるcfcB1変異株はAppppA分解酵素の遺伝子apaHに変異を持つことを明らかにした(西村(昭))。

Research Products

(29 results)

All Other

All Publications (29 results)

[Publications] Feng J.: "New killing system controlled by tow genes located immediately upstream of the mukB gene in E.coli." Mol.Gen.Genet.(in press). (1994)
[Publications] Yamanaka K.: "Characterization of translucent segments observed in an smbA mutant of Escherichia coli." FEMS Microbiol.Lett.(in press). (1994)
[Publications] Yamanaka K.: "Multicopy suppressors,mssA and mssB,of an smbA mutation of Escherichia coli." Mol.Gen.Genet.(in press). (1994)
[Publications] Hiraga S.: "E.coli chromosome partition." Current Opinion in Genetics & Development. 5. 789-801 (1993)
[Publications] Ogawa T.: "Cell cycle-dependent transcription from the gid and mioC promoters of Escherichia coli." J.Bacteriol.(in press). (1994)
[Publications] Masukata H: "Autonomous replication of human chromosomal DNA fragments in human cells." Cell. 4. 1121-1132 (1993)
[Publications] Ohkata K.: "Association of p34^<cdc2>/cycline B complex with microtubules in starfish oocytes." J.Cell Sci.105. 873-881 (1993)
[Publications] Kusubata M.: "cdc2 kinase phosphorylation of desmin at three serine/threonine residues in the amino-terminal head domain." Biochem.Biophys.Res.Commun.190. 927-934 (1993)
[Publications] Ohsumi K.: "Chromosome condensation in Xenopus mitotic extracts without histone Hl." Science. 262. 2033-2035 (1993)
[Publications] Ohta K.: "Microtubule nucleating activity of centrosomes in cell-free extracts from Xenopys eggs:Involvement of phosphorylation and accumulation of pericentriolar material." J.Cell Sci.104. 125-137 (1993)
[Publications] Hisanaga S.: "Tau protein kinase II has a similar characteristic to cdc2 kinase for phoshorylating neurofilament proteins." J.Biol.Chem.268. 15056-15060 (1993)
[Publications] Kataoka T.: "Fully methylated oriC with negative superhelicity forms an oriC-membran complex before initiation of chromosome replication." Biochem.Biophys.Res.Commun.194. 1420-1426 (1993)
[Publications] Tatebayashi K.: "Analysis of integration of DNA fragments by illegitimate recombination in Schizosaccharomyces pombe." Mol.Gen.Genet.(in press). (1994)
[Publications] 加藤潤一: "大腸菌のDNAトポイソメラーゼIV." 細胞工学. 1. 19-26 (1993)
[Publications] Masuda Y.: "Isolation of temperature-sensitive aminoacyl-tRNA synthetase mutants from an Escherichia coli strain harboring the pemK plasmid." Mol.Gen.Genet.238. 169-176 (1993)
[Publications] Tsychimoto S.: "The stable maintenance system pem of plasmid R 100:Degradation of PemI protein may allow PemK protein to inhibit cell growth." J.Bacteriol.174. 4205-4211 (1993)
[Publications] Masuda Y.: "chpA and chpB,Escherichia coli chromosomal homologues of the pem locus responsible for stable maintenance of plasmid R 100." J.Bacteriol.175. 6850-6856 (1993)
[Publications] Higashitani A.: "Osmoregulation of the fatty acid receptor gene fadL in Escherichia coli." Mol.Gen.Genet.240. 339-347 (1993)
[Publications] Tomura A.: "Requirement of phosphatydilglycerol for flagellation of Escherichia coli." FEBS Letters. 329. 287-290 (1993)
[Publications] Ueda T.: "Stabilization of lysozyme by the introduction of Gly-Pro sequence." Protein Engineering. 6. 183-187 (1993)
[Publications] Ogata Y.: "A role of heat shock proteins for homologous recombination in Escherichia coli." Biochem.Biophys.Res.Comm.197. 34-39 (1993)
[Publications] 真木智子: "DNAジャイレースの活性を調節する蛋白質" 細胞工学. 12. 10-18 (1993)
[Publications] Herman C.: "Cell growth and lambda phage development controlled by the same essential Escherichia coli gene,ftsH/hflB." Proc.Natl.Acad.Sci.USA. 90. 10861-10865 (1993)
[Publications] Tomoyasu T.: "Topology and subcellular localization of FtsH protein in Escherichia coli." J.Bacteriol.175. 1352-1357 (1993)
[Publications] Nishitani H.: "Specific chromosomal sites enhancing homologous recombination in Escherichia coli mutants defective in RNaseH." Mol.Gen.Genet.240. 307-314 (1993)
[Publications] Akiyama,Y.: "Involvement of FtsH in protein assembly into and through the membrane.1.Mutation that reduce retention efficiency of a cytoplasmic reporter." J.Biol.Chem.in press. (1994)
[Publications] Tomoyasu,T.: "The Escherichia coli FtsH protein is a prokaryotic member of a protein family of putative ATPases involved in membrane functions,cell cycle control and gene expression." J.Bacteriol.175. 1344-1351 (1993)
[Publications] Masumoto H.: "Properties of CENP-B and its target sequence in satellite DNA,In B.K.Vig(ed),Chromosome Segregation and Aneuploidy." Springer-Verlag,Berlin, 12 (1993)
[Publications] Kishimoto T.: "Control of M-phage by MPF/cyclin-dependent cdc2 kinase.In H.Mohri,M.Takahashi and M.Inagaki(ed),Biology of the Germ Line-Animal and Man." Japan SAcientific Societies Press,Tokyo, 16 (1993)

1993 Fiscal Year Annual Research Report

ゲノムの複製終結と分配制御

Principal Investigator

平賀 壮太 熊本大学, 医学部, 教授 (40027321)

Research Products

[Publications] Feng J.: "New killing system controlled by tow genes located immediately upstream of the mukB gene in E.coli." Mol.Gen.Genet.(in press). (1994)

[Publications] Yamanaka K.: "Characterization of translucent segments observed in an smbA mutant of Escherichia coli." FEMS Microbiol.Lett.(in press). (1994)

[Publications] Yamanaka K.: "Multicopy suppressors,mssA and mssB,of an smbA mutation of Escherichia coli." Mol.Gen.Genet.(in press). (1994)

[Publications] Hiraga S.: "E.coli chromosome partition." Current Opinion in Genetics & Development. 5. 789-801 (1993)

[Publications] Ogawa T.: "Cell cycle-dependent transcription from the gid and mioC promoters of Escherichia coli." J.Bacteriol.(in press). (1994)

[Publications] Masukata H: "Autonomous replication of human chromosomal DNA fragments in human cells." Cell. 4. 1121-1132 (1993)

[Publications] Ohkata K.: "Association of p34^<cdc2>/cycline B complex with microtubules in starfish oocytes." J.Cell Sci.105. 873-881 (1993)

[Publications] Kusubata M.: "cdc2 kinase phosphorylation of desmin at three serine/threonine residues in the amino-terminal head domain." Biochem.Biophys.Res.Commun.190. 927-934 (1993)

[Publications] Ohsumi K.: "Chromosome condensation in Xenopus mitotic extracts without histone Hl." Science. 262. 2033-2035 (1993)

[Publications] Ohta K.: "Microtubule nucleating activity of centrosomes in cell-free extracts from Xenopys eggs:Involvement of phosphorylation and accumulation of pericentriolar material." J.Cell Sci.104. 125-137 (1993)

[Publications] Hisanaga S.: "Tau protein kinase II has a similar characteristic to cdc2 kinase for phoshorylating neurofilament proteins." J.Biol.Chem.268. 15056-15060 (1993)

[Publications] Kataoka T.: "Fully methylated oriC with negative superhelicity forms an oriC-membran complex before initiation of chromosome replication." Biochem.Biophys.Res.Commun.194. 1420-1426 (1993)

[Publications] Tatebayashi K.: "Analysis of integration of DNA fragments by illegitimate recombination in Schizosaccharomyces pombe." Mol.Gen.Genet.(in press). (1994)

[Publications] 加藤潤一: "大腸菌のDNAトポイソメラーゼIV." 細胞工学. 1. 19-26 (1993)

[Publications] Masuda Y.: "Isolation of temperature-sensitive aminoacyl-tRNA synthetase mutants from an Escherichia coli strain harboring the pemK plasmid." Mol.Gen.Genet.238. 169-176 (1993)

[Publications] Tsychimoto S.: "The stable maintenance system pem of plasmid R 100:Degradation of PemI protein may allow PemK protein to inhibit cell growth." J.Bacteriol.174. 4205-4211 (1993)

[Publications] Masuda Y.: "chpA and chpB,Escherichia coli chromosomal homologues of the pem locus responsible for stable maintenance of plasmid R 100." J.Bacteriol.175. 6850-6856 (1993)

[Publications] Higashitani A.: "Osmoregulation of the fatty acid receptor gene fadL in Escherichia coli." Mol.Gen.Genet.240. 339-347 (1993)

[Publications] Tomura A.: "Requirement of phosphatydilglycerol for flagellation of Escherichia coli." FEBS Letters. 329. 287-290 (1993)

[Publications] Ueda T.: "Stabilization of lysozyme by the introduction of Gly-Pro sequence." Protein Engineering. 6. 183-187 (1993)

[Publications] Ogata Y.: "A role of heat shock proteins for homologous recombination in Escherichia coli." Biochem.Biophys.Res.Comm.197. 34-39 (1993)

[Publications] 真木智子: "DNAジャイレースの活性を調節する蛋白質" 細胞工学. 12. 10-18 (1993)

[Publications] Herman C.: "Cell growth and lambda phage development controlled by the same essential Escherichia coli gene,ftsH/hflB." Proc.Natl.Acad.Sci.USA. 90. 10861-10865 (1993)

[Publications] Tomoyasu T.: "Topology and subcellular localization of FtsH protein in Escherichia coli." J.Bacteriol.175. 1352-1357 (1993)

[Publications] Nishitani H.: "Specific chromosomal sites enhancing homologous recombination in Escherichia coli mutants defective in RNaseH." Mol.Gen.Genet.240. 307-314 (1993)

[Publications] Akiyama,Y.: "Involvement of FtsH in protein assembly into and through the membrane.1.Mutation that reduce retention efficiency of a cytoplasmic reporter." J.Biol.Chem.in press. (1994)

[Publications] Tomoyasu,T.: "The Escherichia coli FtsH protein is a prokaryotic member of a protein family of putative ATPases involved in membrane functions,cell cycle control and gene expression." J.Bacteriol.175. 1344-1351 (1993)

[Publications] Masumoto H.: "Properties of CENP-B and its target sequence in satellite DNA,In B.K.Vig(ed),Chromosome Segregation and Aneuploidy." Springer-Verlag,Berlin, 12 (1993)

[Publications] Kishimoto T.: "Control of M-phage by MPF/cyclin-dependent cdc2 kinase.In H.Mohri,M.Takahashi and M.Inagaki(ed),Biology of the Germ Line-Animal and Man." Japan SAcientific Societies Press,Tokyo, 16 (1993)

平賀壮太熊本大学, 医学部, 教授 (40027321)