1999 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 Institution	Kumamoto University
Principal Investigator	平賀壯太熊本大学, 医学部, 教授 (40027321)
Co-Investigator(Kenkyū-buntansha)	和地正明東京工業大学, 生命理工学部, 助教授 (90192822) 大矢禎一東京大学, 大学院・新領域創成科学研究所, 教授 (20183767) 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) 堀内嵩基礎生物学研究所, 教授 (60108644)
Keywords	細胞周期 / DNAメチレーション / 複製開始 / SeqA / DnaA / 染色体分配 / 複製起点
Research Abstract	大腸菌の染色体分配においては、免疫蛍光法、FISH法等の応用により、本年度は次のことが明らかになった。多数のDNAプローブを使って、細胞周期における染色体の動態を解析し、複製準備期には染色体DNAはコンパクトなリング構造をとることを明らかにした。また複製後oriCDNAは互いに対合すること、この対合は約1ラウンドの複製に要する時間の半分であることが明らかになった。半メチル化DNAに結合したSeqA蛋白質が細胞内でクラスターとして存在するが、このクラスターは複製中に2分裂し、細胞中心から2方向に細胞長の1/4,3/4の位置に速やかに移動することがわかった。また、MukB-GFPもSeqAと同様な挙動を示し、姉妹染色体の再構築への関与が示唆された(平賀)。smc欠失株ではoriCの分離・移動に異常が見られる(守家)。新規hda遺伝子はDnaAと相同性をもち、このhda変異では過剰のDNA複製のため致死となる。HdaはDnaAの活性型(DnaA-ATP)から不活性型(DnaA-ADP)への変換に必須な因子である(加藤)。ヒト人工染色体の構造解析から、YACがタンデム連結していることが明らかになった(池野)。 CENP-A、CENP-B、CENP-Cの多重染色により、CEN-AとCEN-Cはドットとして共存し、CENP-Bはその近くに存在していた(杉本)。出芽酵母全てのSPB構成遺伝子のプロモーター領域に転写活性化配列の欠失株は胞子形成過程に異常が見られた(大矢)。細胞壁合成停止時に芽の形成と細胞周期の連携を監視するチェックポイント構造の欠損変異wac1はアクチン類似遺伝子DNAあるARP1の変異である(大矢)。

Research Products
(34 results)

All Other

All Publications (34 results)

[Publications] .Hiraga S.: "Bidirectional migration of SeqA-bound hemimethylated DNA clusters and pairing of oriC copies in Escherichia coli"Genes Cdlls. (in press). (2000)
[Publications] Niki H.: "Dynamic organization of chromosomal DNA in Escherichia coli"Genes Dev. 14. 212-223 (2000)
[Publications] Niki H.: "細菌における分子細胞生物学の新展開-染色体分配と細胞分裂"蛋白質核酸酵素. 45. 153-163 (2000)
[Publications] Kotani S.: "Regulation of APC activity by phosphorylation and regulatory factors"J.Cell Biol.. 146. 791-800 (1999)
[Publications] Ashizawa Y.: "Mechanism of DNA gyrase-mediatedillegitimate recombination:Characterization of Escherichia coli gyrA mutations that confer hyperrecombination phenotype"J.Mol.Biol. 289. 447-458 (1999)
[Publications] Wachi M.: "A murC gene from coryneform bacteria"Appl.Microbiol.Biotechnol.. 51. 223-228 (1999)
[Publications] Wachi M.: "Escherichia coli cafA gene encodes a novel RNase,designated as RNaseG,involved in processing of the 5'end of 16SrRNA"Biochem.Biophys.Res.Commun.. 259. 483-488 (1999)
[Publications] Nagano K.: "fcsA29 mutation in an allele of polA gene of Escherichia coli"Biosci.Biotechnol.Biochem.. 63. 427-429 (1999)
[Publications] Sugimoto K.: "Visualization of prekinetochorelocus on the centromeric region of highly extended chromatin fibers:does kinetochore autoantigen CENP-C constitute a kinetochore organizing center?"Chromosome Res.. 7. 9-19 (1999)
[Publications] Yamazoe,M.: "Complex formation of MukB,MukE and MukF proteins involved in chromosome partitioning in Escherichia coli"EMBO J.. 18. 5873-5884 (1999)
[Publications] Nishito Y.: "Direct metal analyses of Mn^<2+>-dependent and-independent protein phosphatase 2A from human erythrocytes detect zinc and iron only in the Mn^<2+>-independent one."FEBS Lett. 447. 29-33 (1999)
[Publications] Guo L.: "Isolation adn characterization of nevel cold-sensitive dnaA mutants of Escherichia coli"FEBS Micribiol.Lett.. 176. 357-366 (1999)
[Publications] Malatihi K.C.: "Isolation of the murI gene from Brevibacterium lactofermentum ATCC13869 encoding D-glutamate racemase."FEBS Microbiol.Lett.. 175. 193-196 (1999)
[Publications] Lim Y.M.: "Genetic analysis of rolled,which encodes a Drosophila mitogen-activated protein kinase"Genetics. 153. 763-771 (1999)
[Publications] Ali Azam T.: "Growth phase-dependent variation in protein composition of the Escherichia coli nucleoid"J Bacteriol. 181. 6361-6370 (1999)
[Publications] Nihito Y.: "Interconversion of Mn^<2+>-dependent and-independent protein phosphatase 2A from human erythrocytes:role of Zn^<2+> and Fe^<2+> in protein phosphatase 2A"J Biochem(Tokyo). 126. 632-638 (1999)
[Publications] Inoue R.: "Studies on functions of the 63-kDaA-and 74-kDaB'(delta)-regulatory subunits in human erythrocyte protein phosphatase 2A:dissociation and reassociation of the subunits"J Biochem(Tokyo). 126. 1127-1135 (1999)
[Publications] McEwen R.K.: "Complementation analysis in PtdlnsP kinase-deticient yeast mutants demonstrates that Schizosaccharomyces pombe and murine Fab1p homologues are phosphatidylinositol 3-phosphate 5-kinases"J Biol Chem. 274. 33905-33912 (1999)
[Publications] InoueS.B.: "Prenylation of Rho1p is required for activation of yeast1,3-beta-glucan synthase."J Biol Chem. 274. 38119-38124 (1999)
[Publications] Kato J.: "The E.coli homologue of yeast Rer2,a key enzyme of dolichol synthesis,is essential for carrier lipid formation in bacterial cell wall synthesis"J Bacteriol. 181. 2733-2738 (1999)
[Publications] Yamada T.: "Functional domain structure of human heterochromatin protein HP1(Hsalpha):involvement of internal DNA-binding and C-terminal self-association domains in the formation of discrete dots in interphase nuclei"J.Biochem.(Tokyo). 125. 832-837 (1999)
[Publications] Karata K.: "Dissecting the role of a conserved motif(the second region of homology)in the AAA family of ATPases"J.Biol.Chem. 274. 26225-26232 (1999)
[Publications] Sugata N.: "Characterization of a novel kinetochore protein CENP-H"J.Biol.Chem. 274. 27343-27346 (1999)
[Publications] Taki K.: "The SOS response is induced by replication fork blockage at a Ter site located on a pUC-derived plasmid:dependence on the deistance between ori and Ter sites"Mol Gen Genet. 262. 302-309 (1999)
[Publications] Niki H.: "Subcellular localization of plasmids containing the oriC region of the Escherichia coli chromosome,with or without the sopABC partitioning sytem"Mol Microbiol. 34. 498-503 (1999)
[Publications] Sato M.: "The yeast RER2 gene,identified by ER protein localization mutations,encodes cis-prenyltransferase,a key enzyme in the dolichol synthesis"Mol.Cell.Biol.. 19. 471-483 (1999)
[Publications] Ogura T.: "alanced biosynthesis of major membrane components through regulated degradation of the committed enzyme of lipid A biosynthesis by the AAA protease FtsH(HfIB) in Escherichia coli"Mol.Microbiol. 31. 833-844 (1999)
[Publications] Onogi T.: "The assembly and migration of SeqA-Gfp fusion in living cells of Escherichia coli"Mol.Microbiol. 31. 1775-1782 (1999)
[Publications] Kurasawa Y.: "Identification of human APC10/Doc 1 as a subunit of anaphase promoting complex"Oncogene. 18. 5131-5137 (1999)
[Publications] Yamaguchi M.: "Ectopic expression of human p53 inhibits entry into S phase and induces apoptosis in the Drosophila eye imaginal disc"Oncogene. 18. 6767-6775 (1999)
[Publications] Kikyo M.: "An FH domain-containing Bnr1p is a multifunctional protein interacting with a variety of cytoskeletal proteins in Saccharomyces cerevisiae"Oncogene. 18. 7046-7054 (1999)
[Publications] Moriya S.: "Regulation of initiation of Bacillus subtilis chromosome replication"Plasmid. 41. 17-29 (1999)
[Publications] Ohno M.: "間期核の染色体DNA中に存在する三重鎖構造とその機能"蛋白質核酸酵素. 44. 1757-1763 (1999)
[Publications] Araki H.: "間期核の染色体DNA中に存在する三重鎖構造とその機能"蛋白質核酸酵素. 44. 1822-1829 (1999)

1999 Fiscal Year Annual Research Report

染色体分配装置のダイナミクス

Principal Investigator

平賀 壯太 熊本大学, 医学部, 教授 (40027321)

Research Products

[Publications] .Hiraga S.: "Bidirectional migration of SeqA-bound hemimethylated DNA clusters and pairing of oriC copies in Escherichia coli"Genes Cdlls. (in press). (2000)

[Publications] Niki H.: "Dynamic organization of chromosomal DNA in Escherichia coli"Genes Dev. 14. 212-223 (2000)

[Publications] Niki H.: "細菌における分子細胞生物学の新展開-染色体分配と細胞分裂"蛋白質核酸酵素. 45. 153-163 (2000)

[Publications] Kotani S.: "Regulation of APC activity by phosphorylation and regulatory factors"J.Cell Biol.. 146. 791-800 (1999)

[Publications] Ashizawa Y.: "Mechanism of DNA gyrase-mediatedillegitimate recombination:Characterization of Escherichia coli gyrA mutations that confer hyperrecombination phenotype"J.Mol.Biol. 289. 447-458 (1999)

[Publications] Wachi M.: "A murC gene from coryneform bacteria"Appl.Microbiol.Biotechnol.. 51. 223-228 (1999)

[Publications] Wachi M.: "Escherichia coli cafA gene encodes a novel RNase,designated as RNaseG,involved in processing of the 5'end of 16SrRNA"Biochem.Biophys.Res.Commun.. 259. 483-488 (1999)

[Publications] Nagano K.: "fcsA29 mutation in an allele of polA gene of Escherichia coli"Biosci.Biotechnol.Biochem.. 63. 427-429 (1999)

[Publications] Sugimoto K.: "Visualization of prekinetochorelocus on the centromeric region of highly extended chromatin fibers:does kinetochore autoantigen CENP-C constitute a kinetochore organizing center?"Chromosome Res.. 7. 9-19 (1999)

[Publications] Yamazoe,M.: "Complex formation of MukB,MukE and MukF proteins involved in chromosome partitioning in Escherichia coli"EMBO J.. 18. 5873-5884 (1999)

[Publications] Nishito Y.: "Direct metal analyses of Mn^<2+>-dependent and-independent protein phosphatase 2A from human erythrocytes detect zinc and iron only in the Mn^<2+>-independent one."FEBS Lett. 447. 29-33 (1999)

[Publications] Guo L.: "Isolation adn characterization of nevel cold-sensitive dnaA mutants of Escherichia coli"FEBS Micribiol.Lett.. 176. 357-366 (1999)

[Publications] Malatihi K.C.: "Isolation of the murI gene from Brevibacterium lactofermentum ATCC13869 encoding D-glutamate racemase."FEBS Microbiol.Lett.. 175. 193-196 (1999)

[Publications] Lim Y.M.: "Genetic analysis of rolled,which encodes a Drosophila mitogen-activated protein kinase"Genetics. 153. 763-771 (1999)

[Publications] Ali Azam T.: "Growth phase-dependent variation in protein composition of the Escherichia coli nucleoid"J Bacteriol. 181. 6361-6370 (1999)

[Publications] Nihito Y.: "Interconversion of Mn^<2+>-dependent and-independent protein phosphatase 2A from human erythrocytes:role of Zn^<2+> and Fe^<2+> in protein phosphatase 2A"J Biochem(Tokyo). 126. 632-638 (1999)

[Publications] Inoue R.: "Studies on functions of the 63-kDaA-and 74-kDaB'(delta)-regulatory subunits in human erythrocyte protein phosphatase 2A:dissociation and reassociation of the subunits"J Biochem(Tokyo). 126. 1127-1135 (1999)

[Publications] McEwen R.K.: "Complementation analysis in PtdlnsP kinase-deticient yeast mutants demonstrates that Schizosaccharomyces pombe and murine Fab1p homologues are phosphatidylinositol 3-phosphate 5-kinases"J Biol Chem. 274. 33905-33912 (1999)

[Publications] InoueS.B.: "Prenylation of Rho1p is required for activation of yeast1,3-beta-glucan synthase."J Biol Chem. 274. 38119-38124 (1999)

[Publications] Kato J.: "The E.coli homologue of yeast Rer2,a key enzyme of dolichol synthesis,is essential for carrier lipid formation in bacterial cell wall synthesis"J Bacteriol. 181. 2733-2738 (1999)

[Publications] Yamada T.: "Functional domain structure of human heterochromatin protein HP1(Hsalpha):involvement of internal DNA-binding and C-terminal self-association domains in the formation of discrete dots in interphase nuclei"J.Biochem.(Tokyo). 125. 832-837 (1999)

[Publications] Karata K.: "Dissecting the role of a conserved motif(the second region of homology)in the AAA family of ATPases"J.Biol.Chem. 274. 26225-26232 (1999)

[Publications] Sugata N.: "Characterization of a novel kinetochore protein CENP-H"J.Biol.Chem. 274. 27343-27346 (1999)

[Publications] Taki K.: "The SOS response is induced by replication fork blockage at a Ter site located on a pUC-derived plasmid:dependence on the deistance between ori and Ter sites"Mol Gen Genet. 262. 302-309 (1999)

[Publications] Niki H.: "Subcellular localization of plasmids containing the oriC region of the Escherichia coli chromosome,with or without the sopABC partitioning sytem"Mol Microbiol. 34. 498-503 (1999)

[Publications] Sato M.: "The yeast RER2 gene,identified by ER protein localization mutations,encodes cis-prenyltransferase,a key enzyme in the dolichol synthesis"Mol.Cell.Biol.. 19. 471-483 (1999)

[Publications] Ogura T.: "alanced biosynthesis of major membrane components through regulated degradation of the committed enzyme of lipid A biosynthesis by the AAA protease FtsH(HfIB) in Escherichia coli"Mol.Microbiol. 31. 833-844 (1999)

[Publications] Onogi T.: "The assembly and migration of SeqA-Gfp fusion in living cells of Escherichia coli"Mol.Microbiol. 31. 1775-1782 (1999)

[Publications] Kurasawa Y.: "Identification of human APC10/Doc 1 as a subunit of anaphase promoting complex"Oncogene. 18. 5131-5137 (1999)

[Publications] Yamaguchi M.: "Ectopic expression of human p53 inhibits entry into S phase and induces apoptosis in the Drosophila eye imaginal disc"Oncogene. 18. 6767-6775 (1999)

[Publications] Kikyo M.: "An FH domain-containing Bnr1p is a multifunctional protein interacting with a variety of cytoskeletal proteins in Saccharomyces cerevisiae"Oncogene. 18. 7046-7054 (1999)

[Publications] Moriya S.: "Regulation of initiation of Bacillus subtilis chromosome replication"Plasmid. 41. 17-29 (1999)

[Publications] Ohno M.: "間期核の染色体DNA中に存在する三重鎖構造とその機能"蛋白質核酸酵素. 44. 1757-1763 (1999)

[Publications] Araki H.: "間期核の染色体DNA中に存在する三重鎖構造とその機能"蛋白質核酸酵素. 44. 1822-1829 (1999)

平賀壯太熊本大学, 医学部, 教授 (40027321)