1998 年度実績報告書

染色体分配装置のダイナミクス

研究課題

研究課題/領域番号	08277104
研究種目	特定領域研究(A)
研究機関	熊本大学
研究代表者	平賀莊太熊本大学, 医学部, 教授 (40027321)
研究分担者	加藤潤一東京大学, 医科学研究所, 助手 (10194820) 堀内嵩基礎生物学研究所, 教授 (60108644) 大矢禎一東京大学大学院, 理学系研究科, 助教授 (20183767) 和地正明東京工業大学, 生命理工学部, 助教授 (90192822) 小椋光熊本大学, 医学部, 助教授 (00158825)
キーワード	大腸菌染色体 / 免疫蛍光顕微鏡法 / oriCDNA / 細胞周期 / ミニFプラスミドDNA / 枯草菌 / 人工ミニ染色体 / ヒトセントロメア
研究概要	1. 染色体上の多数のDNAプローブを使ったFISH法の解析によって、大腸菌の染色体DNAは細菌内で太い環状になっていることがわかった。免疫蛍光顕微鏡法で染色体複製調節に関与するSeqA蛋白の細胞内分布を種々の高温感受性複製変異株において解析し、その結果を総合して新しいモデルを提出した(平賀)。 2. ミニFプラスミドの安定保持ができなくなった大腸菌の変異株の解析から、DnaAと相同性を持つ新規遺伝子hdaを見つけた(加藤)。 3. 枯草菌smc欠損変異株では染色体分離が異常になると共に複製開始領域の細胞内分布がランダムになる。Smcの大量発現系を作りsmcの精製を試みている(守屋)。 4. muk変異株を用いたスクリーニングによって、染色体の分配過程を特異的に阻害する多数の化合物を発見した。その中の多くはDNAジャイレースやトポイソメラーゼを阻害した(和地)。 5. 免疫共沈実験によりヒトCENP-AとCENP-Bとがヌクレオゾーム構造レベルで極めて近傍に存在し、DNAを介して複合体を形成していることがわかった(依田)。 6. ヒトへテロクノマチン蛋白質HP1とGFPの融合蛋白質は細胞内にヘテロクロマチン様「ドット」として局在し、また中央体にも局在した。CENP-C、CENP-B、アルフォイドDNAのゲノム上での視覚化に成功した(杉本)。 7. 導入したYACがタンデムに巨大化しヒトミニ染色体を形成するメカニズムについて解析し、導入したYACの末端部にまずエピゾーマルDNA断片が結合し、その後この単位が増幅してYACの数十倍の大きさの人工染色体が形成されることが明らかになった(池野)。 8. 細胞内の全DnaA蛋白質の約60%が結合するdatA DNA領域が大腸菌の染色体複製開始の調節に関与することを明らかにした(小川)。

研究成果
(30件)

すべてその他

すべて文献書誌 (30件)

[文献書誌] Niki,H.: "Polar of the replication origin and terminus in Escherichia coli nucleoids during chromosome partitioning." Genus&Development. 12. 1036-1045 (1998)
[文献書誌] Hirano,M.: "Autoregulation of the partition genus of mini-F plasmid and the intracellilar localization of their products in Escherichia coli." Mol.Gen.Genet.257. 392-403 (1998)
[文献書誌] Hiraga,S.: "Cell cycle-dependent duplication and bidirectional migration of SeqA-associated DNA-protein comlexes in E.coli." Mol.Cell.1. 381-387 (1998)
[文献書誌] Okano,H.: "Importance of Phenylalanine Residues of yeast Calmodulin for Target Binding and Activation." J.Biol.Chem.273. 26375-26382 (1998)
[文献書誌] Sekiya,-K.M.: "Identification of Functional Connections between Calmodulin and the yeast Actin Cytoskeleton." Genetics. 150. 43-58 (1998)
[文献書誌] Homma,K.: "Phosphatidylinositol-4-Phosphate 5-Kinase Localized on the plasma Membrane Is Essential for Yeast Cell Morphogenesis." J.Biol.Chem.273. 15779-15786 (1998)
[文献書誌] Ukai,H.: "ftsE(Ts)affect translocation of K^+ -pump proteins into the membrane of Escherichia coli." J.Bacteriol.180. 3663-3670 (1998)
[文献書誌] Nishimura,A: "Timing of cell division:Ap4A as the signal." TiBS. 23. 157-159 (1998)
[文献書誌] Heo,S-j.: "The RHC21 gene of budding yeast,a homologue of the fission yeast rad21^+ gene,is essential for chromosome segregation." Mol.Gen.Genet.257. 149-156 (1998)
[文献書誌] Tatebayashi,K: "Isolation of a Schizosaccharomyces pombe rad21st mutant that is aberrant in chromosome segregation,microtubule function,DNA repair and sensetive to hydroxyurea:possible involvement of Rad21 in ubiquitinmediated proteolysis." Genetics. 148. 49-57 (1998)
[文献書誌] Yamagata,K.: "Bloom's and Werner's syndrime genes suppress hyperrecombination in yeast sgs1 mutant:implication for genomic instability in human diseases." Proc.Natl.Acad.Sci.USA. 95. 8733-8738 (1998)
[文献書誌] Shanado,Y.: "Escherichia coli HU protein suppresses DNA gyrase-mediated illegitimate recombination and SOS induction." Genes to Cells. 3. 511-250 (1998)
[文献書誌] Caplin B.E.: "Amino Acid Residues That Define Both the Isoprenoid and CAAX Preferences of the Saccharomyces cerevisiae Protein Farnesyltransferase.Creating the perfect farnesyltransferase." J.Biol.Chem.273. 9472-9479 (1998)
[文献書誌] Helliwell,S.B.: "The RH01 effector PKC1,but not BNL1,mediates signalling from TOR2 to the actin cytoskeleton." Curr.Biol.22. 1211-1214 (1998)
[文献書誌] Kijima,N.: "Induction of only limited elongation instead of filamentation by inhibition of cell division in Corynebacterium glutamicum." Appl.Microbiol.Biotechnol.50. 227-232 (1998)
[文献書誌] 和地正明: "コリネ型細菌の細胞複製機構の解析" バイオサイエンスとインダストリー. 56. 101-103 (1998)
[文献書誌] 和地正明: "大腸菌の細胞周期と核様体タンパク質" バイオサイエンスとインダストリー. 56. 229-232 (1998)
[文献書誌] Kobayashi,T.: "Expansion and contraction of ribosomal DNA repeats in Saccharomyces cerevisiae:requirement of replication fork blocking(Fobl)protein and the role of RNA polymerase I." Gene&Development. 12. 3821-3830 (1998)
[文献書誌] Kitagawa,R.: "Negative control of replication initiation by a novel chromosomal locus exhibiting exceptional affinity for Escherichia coli DnaA protein." Genes&Development. 12. 3032-3043 (1998)
[文献書誌] Mizushima,T.: "Site-directed mutational analysis for the ATP binding of DnaA prtein.Functions of two conserved amino acids(Lys-178 and Asp-235)located in the ATP-binding domain of DnaA protein in vitro and in vivo." J.Biol.Chem.273. 20847-20851 (1998)
[文献書誌] Hashimoto,Y.: "Construction of a yeast expression system with positive-selection in the absence of a specific phenotype." Gen. 207. 167-170 (1998)
[文献書誌] Yoda,K.: "In vitro assembly of CENP-B/alpha-satellite DNA/core histon comlex:CENP-B causes nucleosome positioning." Genes to Cells. 3. 533-548 (1998)
[文献書誌] Moriya,S.: "A Bacilus subtilis gene-encoding protein homologous to eukaryotic SMC motor protein is necessary for chromosome partition." Mol.Microbiol.29. 179-187 (1998)
[文献書誌] Nanamiya,H.: "ClpC regulates the fate of a sporulation initiation sigma factor,s H protein,in Bacillus subtilis at elevated temperatures." Mol.Microbiol.29. 505-513 (1998)
[文献書誌] Sugimoto,K: "Nucleotide specificity at the boundary and size requirement of the target sites recognized by human centromere protein B(CENP-B)in vitro." Chromosome Res.6. 133-140 (1998)
[文献書誌] Ikeno,M.: "Construction of YAC-based mammalianatificial chromosomes." Nature Biotechnolozy. 16. 431-439 (1998)
[文献書誌] Tomoyasu,T.: "Levels of DnaK and DnaJ provide tight control of heat shock gene expression and protein repair in Escherichia coli." Mol.Microbiol.30. 567-581 (1998)
[文献書誌] Ogura,T.: "Balanced biosynthesis of major membrane components through regulated degradation of the committed enzyme of lipid A biosynthesis by the AAA protease FisH(HfIB)in Escherichia coli." Mol.Microbiol.31. 833-844 (1998)
[文献書誌] Akiyama,Y.: "Roles of the periplamic domain of Escherichia coli FtsH(HfIB),in protein interactions and activity modulation." J.Biol.Chem.28. 22326-22333 (1998)
[文献書誌] Tatsuta,T.: "Heat shock regulation in the FtsH null mutant of Escherichia coli:dissection of stability and activity control mechanisms of σ^<32> in vivo." Mol.Microbiol.30. 583-593 (1998)

1998 年度 実績報告書

染色体分配装置のダイナミクス

研究代表者

平賀 莊太 熊本大学, 医学部, 教授 (40027321)

研究成果

[文献書誌] Niki,H.: "Polar of the replication origin and terminus in Escherichia coli nucleoids during chromosome partitioning." Genus&Development. 12. 1036-1045 (1998)

[文献書誌] Hirano,M.: "Autoregulation of the partition genus of mini-F plasmid and the intracellilar localization of their products in Escherichia coli." Mol.Gen.Genet.257. 392-403 (1998)

[文献書誌] Hiraga,S.: "Cell cycle-dependent duplication and bidirectional migration of SeqA-associated DNA-protein comlexes in E.coli." Mol.Cell.1. 381-387 (1998)

[文献書誌] Okano,H.: "Importance of Phenylalanine Residues of yeast Calmodulin for Target Binding and Activation." J.Biol.Chem.273. 26375-26382 (1998)

[文献書誌] Sekiya,-K.M.: "Identification of Functional Connections between Calmodulin and the yeast Actin Cytoskeleton." Genetics. 150. 43-58 (1998)

[文献書誌] Homma,K.: "Phosphatidylinositol-4-Phosphate 5-Kinase Localized on the plasma Membrane Is Essential for Yeast Cell Morphogenesis." J.Biol.Chem.273. 15779-15786 (1998)

[文献書誌] Ukai,H.: "ftsE(Ts)affect translocation of K^+ -pump proteins into the membrane of Escherichia coli." J.Bacteriol.180. 3663-3670 (1998)

[文献書誌] Nishimura,A: "Timing of cell division:Ap4A as the signal." TiBS. 23. 157-159 (1998)

[文献書誌] Heo,S-j.: "The RHC21 gene of budding yeast,a homologue of the fission yeast rad21^+ gene,is essential for chromosome segregation." Mol.Gen.Genet.257. 149-156 (1998)

[文献書誌] Tatebayashi,K: "Isolation of a Schizosaccharomyces pombe rad21st mutant that is aberrant in chromosome segregation,microtubule function,DNA repair and sensetive to hydroxyurea:possible involvement of Rad21 in ubiquitinmediated proteolysis." Genetics. 148. 49-57 (1998)

[文献書誌] Yamagata,K.: "Bloom's and Werner's syndrime genes suppress hyperrecombination in yeast sgs1 mutant:implication for genomic instability in human diseases." Proc.Natl.Acad.Sci.USA. 95. 8733-8738 (1998)

[文献書誌] Shanado,Y.: "Escherichia coli HU protein suppresses DNA gyrase-mediated illegitimate recombination and SOS induction." Genes to Cells. 3. 511-250 (1998)

[文献書誌] Caplin B.E.: "Amino Acid Residues That Define Both the Isoprenoid and CAAX Preferences of the Saccharomyces cerevisiae Protein Farnesyltransferase.Creating the perfect farnesyltransferase." J.Biol.Chem.273. 9472-9479 (1998)

[文献書誌] Helliwell,S.B.: "The RH01 effector PKC1,but not BNL1,mediates signalling from TOR2 to the actin cytoskeleton." Curr.Biol.22. 1211-1214 (1998)

[文献書誌] Kijima,N.: "Induction of only limited elongation instead of filamentation by inhibition of cell division in Corynebacterium glutamicum." Appl.Microbiol.Biotechnol.50. 227-232 (1998)

[文献書誌] 和地正明: "コリネ型細菌の細胞複製機構の解析" バイオサイエンスとインダストリー. 56. 101-103 (1998)

[文献書誌] 和地正明: "大腸菌の細胞周期と核様体タンパク質" バイオサイエンスとインダストリー. 56. 229-232 (1998)

[文献書誌] Kobayashi,T.: "Expansion and contraction of ribosomal DNA repeats in Saccharomyces cerevisiae:requirement of replication fork blocking(Fobl)protein and the role of RNA polymerase I." Gene&Development. 12. 3821-3830 (1998)

[文献書誌] Kitagawa,R.: "Negative control of replication initiation by a novel chromosomal locus exhibiting exceptional affinity for Escherichia coli DnaA protein." Genes&Development. 12. 3032-3043 (1998)

[文献書誌] Mizushima,T.: "Site-directed mutational analysis for the ATP binding of DnaA prtein.Functions of two conserved amino acids(Lys-178 and Asp-235)located in the ATP-binding domain of DnaA protein in vitro and in vivo." J.Biol.Chem.273. 20847-20851 (1998)

[文献書誌] Hashimoto,Y.: "Construction of a yeast expression system with positive-selection in the absence of a specific phenotype." Gen. 207. 167-170 (1998)

[文献書誌] Yoda,K.: "In vitro assembly of CENP-B/alpha-satellite DNA/core histon comlex:CENP-B causes nucleosome positioning." Genes to Cells. 3. 533-548 (1998)

[文献書誌] Moriya,S.: "A Bacilus subtilis gene-encoding protein homologous to eukaryotic SMC motor protein is necessary for chromosome partition." Mol.Microbiol.29. 179-187 (1998)

[文献書誌] Nanamiya,H.: "ClpC regulates the fate of a sporulation initiation sigma factor,s H protein,in Bacillus subtilis at elevated temperatures." Mol.Microbiol.29. 505-513 (1998)

[文献書誌] Sugimoto,K: "Nucleotide specificity at the boundary and size requirement of the target sites recognized by human centromere protein B(CENP-B)in vitro." Chromosome Res.6. 133-140 (1998)

[文献書誌] Ikeno,M.: "Construction of YAC-based mammalianatificial chromosomes." Nature Biotechnolozy. 16. 431-439 (1998)

[文献書誌] Tomoyasu,T.: "Levels of DnaK and DnaJ provide tight control of heat shock gene expression and protein repair in Escherichia coli." Mol.Microbiol.30. 567-581 (1998)

[文献書誌] Ogura,T.: "Balanced biosynthesis of major membrane components through regulated degradation of the committed enzyme of lipid A biosynthesis by the AAA protease FisH(HfIB)in Escherichia coli." Mol.Microbiol.31. 833-844 (1998)

[文献書誌] Akiyama,Y.: "Roles of the periplamic domain of Escherichia coli FtsH(HfIB),in protein interactions and activity modulation." J.Biol.Chem.28. 22326-22333 (1998)

[文献書誌] Tatsuta,T.: "Heat shock regulation in the FtsH null mutant of Escherichia coli:dissection of stability and activity control mechanisms of σ^<32> in vivo." Mol.Microbiol.30. 583-593 (1998)

1998 年度実績報告書

平賀莊太熊本大学, 医学部, 教授 (40027321)