1993 年度実績報告書

G1/S移行期の制御機構

研究課題

研究課題/領域番号	05269101
研究機関	東京大学
研究代表者	東江昭夫東京大学, 大学院・理学系研究科, 教授 (90029249)
研究分担者	田中一馬神戸大学, 医学部, 助手 (60188290) 南康博大阪大学, 細胞生体工学センター, 助手 (70229772) 荒木弘之大阪大学, 微生物病研究所, 助教授 (20151160) 秋山徹大阪大学, 微生物病研究所, 助教授 (70150745) 宮川都吉広島大学, 工学部, 教授 (10116676)
キーワード	細胞周期 / 信号伝達 / 出芽酵母 / 線維芽細胞 / IL-2 / 低分子量G蛋白質 / DNA複製
研究概要	出芽酵母のM期終了に新規の低分子量G蛋白質が関与することを見いだした。このG蛋白質遺伝子TEM1は必須遺伝子で、M期終了に関わる蛋白質リン酸化酵素Cdc15の作用を活性化しMPFを不活化すると考えている(東江)。信号伝達系とG1/S移行期の接点として、出芽酵母のCa^<++>依存性プロテインホスファターゼ(PP2B)を介するシグナル伝達機構を調べ、PP2Bはイオンストレス下に細胞内イオン恒常性を維持するのに必要であった。また、Ca^<++>によるPP2Bの恒常的活性化は致死的なことを示す結果を得た(宮川)。機能的高親和性IL-2受容体を線維芽細胞に再構成し、その機能解析を行った。その結果、IL-2受容体は、IL-2刺激に伴うMAPキナーゼのチロシンリン酸化昂進、および核内プロトオンコジーン(c-fos,c-jun、c-myc遺伝子)の発現誘導などの生物学的変化を、リンパ球の場合と同様に引き起こすことができるにもかかわらず、G1期からS期への進行を誘導できないことが判った(南)。RB結合蛋白質RAXがRB蛋白質と協同してc-fos遺伝子の転写を抑制する活性を持つことを明らかにした。また、RAX蛋白質が骨肉腫由来Saos2細胞や膀胱癌由来HTB9細胞の増殖を強く抑制することを見いだした。これらの結果から、RAXは負の転写因子と考えられた。一方、WT1蛋白質が細胞周期のG1からS期への移行を阻害する活性を持つことを見いだした(秋山)。出芽酵母の低分子量G蛋白質の一つRho1が欠損すると、細胞は小さな芽を出した状態で増殖を停止する。免疫蛍光抗体法によりRho1蛋白質は芽の生長点に局在することが明らかになった(田中)。出芽酵母のDNAポリメラーゼIIの変異体、温度感受性po12-18株から高温で増殖できる劣性復帰変異株を分離し5グループに分類した。その内の一つは低温感受性でPolyA binding proteinに類似の新規RNP遺伝子の変異によることが判った。もう一つのグループはDNAポリメラーゼIの変異も抑圧し、単独で弱い高温感受性を示した(荒木)。

研究成果

(28件)

すべてその他

すべて文献書誌 (28件)

[文献書誌] Yoko-o,T.et al.: "The putative phosphoinositide-specific phospholipase C gene,PLC1,of the yeast Saccharomyces cerevisiae is important for cell growth." Proc.Natl.Acad.Sci.USA. 90. 1804-1808 (1993)
[文献書誌] Toh-e,A.et al.: "Three yeast genes,PIR1,PIR2 and PIR3,containing internal tandem repeats,are related to each other,and PIR1 and PIR2 are required for tolerance to heat shock." Yeast. 9. 481-494 (1993)
[文献書誌] Shirayama,M.et al.: "MISI3,a multicopy suppressor of mutants hyperactivated in the RAS-cAMP pathway,encodes a novel HSP70 protein of Saccharomyces cerevisiae." Mol.Gen.Genet.240. 323-332 (1993)
[文献書誌] Fujimura,K.et al.: "A dominant interfering mutation in RAS1 of Saccharomyces cerevisiae." Mol.Gen.Genet.241. 280-286 (1993)
[文献書誌] Nakafuku,M.et al.: "Suppression of oncogenic RAS by mutant neurofibromatosis type I genes with single amino acid substitutions." Proc.Natl.Acad.Sci.USA. 91. 6706-6710 (1993)
[文献書誌] Yoshida,M.et al.: "Cloning and sequencing of the NES24 gene of Saccharomyces cerevisiae." Yeast. 10(in press). (1994)
[文献書誌] Shirayama,M.et al.: "Identification of CDC25 family gene,MSI2/LTE1,as a multicopy suppressor of iral." Yeast. 10(in press). (1994)
[文献書誌] Fujimura,M.et al.: "The Saccharomyces cerevisiae MSI4 gene encodes the yeast counterpart of component A of Rab geranylgeranyl transferease." J.Biol.Chem.269(in press). (1994)
[文献書誌] Uesono,Y.et al.: "The MCS1/SSD1/SRK1/SSL1 gene is involved in stable maintenance of the chromosome in yeast." Gene. (in press). (1994)
[文献書誌] Ikeda,E.et al.: "YGE1 is a yeast homologue of Escherichia coli grpE and required for maintenance of mitochondrial functions." FEBS Letters. (in press). (1994)
[文献書誌] Kominami,K.et al.: "Characterization of the function of the NIN1 gene product of Saccharomyces cerevisiae." Exp.Cell Res.211(in press). (1994)
[文献書誌] Nakamura,T.et al.: "Calcineurin-mediated,FK506-sensitive regulation of intracellular ions in yeast is an important determinant for adaptation to high salt stress conditions." EMBO J.12. 4063-4071 (1993)
[文献書誌] Hirata,D.et al.: "Stress-induced transcriptional activation mediated by YAP1 and YAP2 genes that encode the Jun family of transcriptional activation in Saccharomyces cerevisiae." Mol.Gen.Genet.(in press). (199)
[文献書誌] Mukai,H.et al.: "Isolation and characterization of SSE1 and SSE2,new members of the yeast HSP70 multigene family." Gene. 132. 57-66 (1993)
[文献書誌] Hiraga,K.et al.: "Identification and characterization of nuclear calmodulin-binding proteins of Saccharomyces cerevisiae." Biochim.Biophys.Acta. 1177. 25-30 (1993)
[文献書誌] Hiraga,K.et al.: "Cloning and characterization of elongation factor EF-1b homologue of S.cerevisiae:EF-1b is essential for growth." FEBS Letters. 316. 165-169 (1993)
[文献書誌] Irie,K.et al.: "A gene,SMP2,involved in plasmid maintenance and respiration in Saccharomyces cerevisiae encodes a highly chraged protein." Mol.Gen.Genet.236. 283-288 (1993)
[文献書誌] Araki,H.et al.: "A specific host factor binds at a cis-acting locus silent in transcription but functional for stability control of a yeast plamsmid pSR1." Mol.Gen.Genet.238. 120-128 (1993)
[文献書誌] Lee,K.S.et al.: "A yeast mitogen-activated protein kinase homolog (Mpklp) mediates signalling by protein kinase C." Mol.Cell.Biol.13. 3067-3075 (1993)
[文献書誌] Irie,K.et al.: "MKK1 and MKK2,which encode Saccharomyces cerevisiae mitogen-activated protein kinase-kinase homologs,function in the pathway mediated by protein kinase C." Mol.Cell.Biol.13. 3076-3083 (1993)
[文献書誌] Shimizu,K.et al.: "Suppression of bacteriophage T7 ssb mutation with host ssb." J.Mol.Biol.234. 926-931 (1993)
[文献書誌] Minami,Y.et al.: "Association of p56^<lck> with IL-2 receptor B chain is critical for the IL-2-induced activation of p56^<lck>." EMBO J.12. 759-768 (1993)
[文献書誌] Kobayashi,N.et al.: "Functional coupling of the src-family protein tyrosine kinases p59^<fyn> and p53/p56^<fyn> with the interleukin 2 receptor:Implications for redundancy and pleiotropism in cytokine signal transduction." Proc.Natl.Acad.Sci.USA. 90. 4201-4205 (1993)
[文献書誌] Minami,Y.et al.: "The IL-2 receptor complex:Its structure,function,and target genes." Ann.Rev.Immunol.11. 245-267 (1993)
[文献書誌] Kobayashi,N.et al.: "Cloning and sequencing of the cDNA encoding a mouse IL-2 receptor γ." Gene. 130. 303-304 (1993)
[文献書誌] Taniguchi,T.et al.: "The IL-2/IL-2 receptor system:A current overview." Cell. 73. 5-8 (1993)
[文献書誌] Kono,T.et al.: "The interleukin-2 receptor mediated intracellular signal transduction." Seminars in Immunology. 5. 299-307 (1993)
[文献書誌] Samelson,L.E.et al.: "Molecular Mechanisms of Immunological Self-Recognition" Academic Press,Inc., 14 (1993)

1993 年度 実績報告書

G1/S移行期の制御機構

研究代表者

東江 昭夫 東京大学, 大学院・理学系研究科, 教授 (90029249)

研究成果

[文献書誌] Yoko-o,T.et al.: "The putative phosphoinositide-specific phospholipase C gene,PLC1,of the yeast Saccharomyces cerevisiae is important for cell growth." Proc.Natl.Acad.Sci.USA. 90. 1804-1808 (1993)

[文献書誌] Toh-e,A.et al.: "Three yeast genes,PIR1,PIR2 and PIR3,containing internal tandem repeats,are related to each other,and PIR1 and PIR2 are required for tolerance to heat shock." Yeast. 9. 481-494 (1993)

[文献書誌] Shirayama,M.et al.: "MISI3,a multicopy suppressor of mutants hyperactivated in the RAS-cAMP pathway,encodes a novel HSP70 protein of Saccharomyces cerevisiae." Mol.Gen.Genet.240. 323-332 (1993)

[文献書誌] Fujimura,K.et al.: "A dominant interfering mutation in RAS1 of Saccharomyces cerevisiae." Mol.Gen.Genet.241. 280-286 (1993)

[文献書誌] Nakafuku,M.et al.: "Suppression of oncogenic RAS by mutant neurofibromatosis type I genes with single amino acid substitutions." Proc.Natl.Acad.Sci.USA. 91. 6706-6710 (1993)

[文献書誌] Yoshida,M.et al.: "Cloning and sequencing of the NES24 gene of Saccharomyces cerevisiae." Yeast. 10(in press). (1994)

[文献書誌] Shirayama,M.et al.: "Identification of CDC25 family gene,MSI2/LTE1,as a multicopy suppressor of iral." Yeast. 10(in press). (1994)

[文献書誌] Fujimura,M.et al.: "The Saccharomyces cerevisiae MSI4 gene encodes the yeast counterpart of component A of Rab geranylgeranyl transferease." J.Biol.Chem.269(in press). (1994)

[文献書誌] Uesono,Y.et al.: "The MCS1/SSD1/SRK1/SSL1 gene is involved in stable maintenance of the chromosome in yeast." Gene. (in press). (1994)

[文献書誌] Ikeda,E.et al.: "YGE1 is a yeast homologue of Escherichia coli grpE and required for maintenance of mitochondrial functions." FEBS Letters. (in press). (1994)

[文献書誌] Kominami,K.et al.: "Characterization of the function of the NIN1 gene product of Saccharomyces cerevisiae." Exp.Cell Res.211(in press). (1994)

[文献書誌] Nakamura,T.et al.: "Calcineurin-mediated,FK506-sensitive regulation of intracellular ions in yeast is an important determinant for adaptation to high salt stress conditions." EMBO J.12. 4063-4071 (1993)

[文献書誌] Hirata,D.et al.: "Stress-induced transcriptional activation mediated by YAP1 and YAP2 genes that encode the Jun family of transcriptional activation in Saccharomyces cerevisiae." Mol.Gen.Genet.(in press). (199)

[文献書誌] Mukai,H.et al.: "Isolation and characterization of SSE1 and SSE2,new members of the yeast HSP70 multigene family." Gene. 132. 57-66 (1993)

[文献書誌] Hiraga,K.et al.: "Identification and characterization of nuclear calmodulin-binding proteins of Saccharomyces cerevisiae." Biochim.Biophys.Acta. 1177. 25-30 (1993)

[文献書誌] Hiraga,K.et al.: "Cloning and characterization of elongation factor EF-1b homologue of S.cerevisiae:EF-1b is essential for growth." FEBS Letters. 316. 165-169 (1993)

[文献書誌] Irie,K.et al.: "A gene,SMP2,involved in plasmid maintenance and respiration in Saccharomyces cerevisiae encodes a highly chraged protein." Mol.Gen.Genet.236. 283-288 (1993)

[文献書誌] Araki,H.et al.: "A specific host factor binds at a cis-acting locus silent in transcription but functional for stability control of a yeast plamsmid pSR1." Mol.Gen.Genet.238. 120-128 (1993)

[文献書誌] Lee,K.S.et al.: "A yeast mitogen-activated protein kinase homolog (Mpklp) mediates signalling by protein kinase C." Mol.Cell.Biol.13. 3067-3075 (1993)

[文献書誌] Irie,K.et al.: "MKK1 and MKK2,which encode Saccharomyces cerevisiae mitogen-activated protein kinase-kinase homologs,function in the pathway mediated by protein kinase C." Mol.Cell.Biol.13. 3076-3083 (1993)

[文献書誌] Shimizu,K.et al.: "Suppression of bacteriophage T7 ssb mutation with host ssb." J.Mol.Biol.234. 926-931 (1993)

[文献書誌] Minami,Y.et al.: "Association of p56^<lck> with IL-2 receptor B chain is critical for the IL-2-induced activation of p56^<lck>." EMBO J.12. 759-768 (1993)

[文献書誌] Kobayashi,N.et al.: "Functional coupling of the src-family protein tyrosine kinases p59^<fyn> and p53/p56^<fyn> with the interleukin 2 receptor:Implications for redundancy and pleiotropism in cytokine signal transduction." Proc.Natl.Acad.Sci.USA. 90. 4201-4205 (1993)

[文献書誌] Minami,Y.et al.: "The IL-2 receptor complex:Its structure,function,and target genes." Ann.Rev.Immunol.11. 245-267 (1993)

[文献書誌] Kobayashi,N.et al.: "Cloning and sequencing of the cDNA encoding a mouse IL-2 receptor γ." Gene. 130. 303-304 (1993)

[文献書誌] Taniguchi,T.et al.: "The IL-2/IL-2 receptor system:A current overview." Cell. 73. 5-8 (1993)

[文献書誌] Kono,T.et al.: "The interleukin-2 receptor mediated intracellular signal transduction." Seminars in Immunology. 5. 299-307 (1993)

[文献書誌] Samelson,L.E.et al.: "Molecular Mechanisms of Immunological Self-Recognition" Academic Press,Inc., 14 (1993)

1993 年度実績報告書

東江昭夫東京大学, 大学院・理学系研究科, 教授 (90029249)