1998 Fiscal Year Annual Research Report

染色体分配装置のダイナミクス

Research Project

Project/Area Number	08277104
Research Category	Grant-in-Aid for Scientific Research on Priority Areas (A)
Research Institution	Kumamoto University
Principal Investigator	平賀莊太熊本大学, 医学部, 教授 (40027321)
Co-Investigator(Kenkyū-buntansha)	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)
Keywords	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
Research Abstract	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)。

Research Products
(30 results)

All Other

All Publications (30 results)

[Publications] Niki,H.: "Polar of the replication origin and terminus in Escherichia coli nucleoids during chromosome partitioning." Genus&Development. 12. 1036-1045 (1998)
[Publications] 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)
[Publications] Hiraga,S.: "Cell cycle-dependent duplication and bidirectional migration of SeqA-associated DNA-protein comlexes in E.coli." Mol.Cell.1. 381-387 (1998)
[Publications] Okano,H.: "Importance of Phenylalanine Residues of yeast Calmodulin for Target Binding and Activation." J.Biol.Chem.273. 26375-26382 (1998)
[Publications] Sekiya,-K.M.: "Identification of Functional Connections between Calmodulin and the yeast Actin Cytoskeleton." Genetics. 150. 43-58 (1998)
[Publications] 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)
[Publications] Ukai,H.: "ftsE(Ts)affect translocation of K^+ -pump proteins into the membrane of Escherichia coli." J.Bacteriol.180. 3663-3670 (1998)
[Publications] Nishimura,A: "Timing of cell division:Ap4A as the signal." TiBS. 23. 157-159 (1998)
[Publications] 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)
[Publications] 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)
[Publications] 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)
[Publications] Shanado,Y.: "Escherichia coli HU protein suppresses DNA gyrase-mediated illegitimate recombination and SOS induction." Genes to Cells. 3. 511-250 (1998)
[Publications] 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)
[Publications] Helliwell,S.B.: "The RH01 effector PKC1,but not BNL1,mediates signalling from TOR2 to the actin cytoskeleton." Curr.Biol.22. 1211-1214 (1998)
[Publications] 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)
[Publications] 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)
[Publications] 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)
[Publications] 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)
[Publications] 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)
[Publications] 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)
[Publications] Hashimoto,Y.: "Construction of a yeast expression system with positive-selection in the absence of a specific phenotype." Gen. 207. 167-170 (1998)
[Publications] 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)
[Publications] 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)
[Publications] 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)
[Publications] 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)
[Publications] Ikeno,M.: "Construction of YAC-based mammalianatificial chromosomes." Nature Biotechnolozy. 16. 431-439 (1998)
[Publications] 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)
[Publications] 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)
[Publications] 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)
[Publications] 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 Fiscal Year Annual Research Report

染色体分配装置のダイナミクス

Principal Investigator

平賀 莊太 熊本大学, 医学部, 教授 (40027321)

Research Products

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

[Publications] 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)

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

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

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

[Publications] 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)

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

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

[Publications] 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)

[Publications] 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)

[Publications] 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)

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

[Publications] 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)

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

[Publications] 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)

[Publications] 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)

[Publications] 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)

[Publications] 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)

[Publications] 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)

[Publications] 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)

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

[Publications] 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)

[Publications] 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)

[Publications] 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)

[Publications] 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)

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

[Publications] 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)

[Publications] 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)

[Publications] 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)

[Publications] 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)

平賀莊太熊本大学, 医学部, 教授 (40027321)