1996 年度実績報告書

挿入突然変異体による粘菌細胞の分化と形態形成の解析

研究課題

研究課題/領域番号	07308052
研究機関	岡崎国立共同研究機構
研究代表者	竹内郁夫岡崎国立共同研究機構, 機構長 (90025239)
研究分担者	前田ミネ子大阪大学, 大学院・理学研究科, 助手 (70029700) 田仲可昌筑波大学, 生物科学系, 教授 (80091908) 前田靖男東北大学, 大学院・理学研究科, 教授 (50025417) 落合廣北海道大学, 大学院・理学研究科, 教授 (10002122) 須藤和夫東京大学, 教養学部, 助教授 (20111453)
キーワード	細胞性粘菌 / 挿入突然変異体 / 遺伝子工学 / 形態形成 / 発生 / 細胞分化 / Dictyostelium / シグナル伝達
研究概要	竹内は粘菌のcDNAプロジェクトを昨年秋からスタートさせた。また,ガラスキャピラリー内に吸い込んだ粘菌細胞では、正常発生の場合と異なり、予定柄・予定胞子細胞が一定の比率で分化することはないことを明らかにした。形態形成の分子機構須藤は、挿入突然変異法により生じた、集合期以降の発生が起こらない変異体の遺伝子を解析して酵母のSSN6遺伝子の相同体を分離し、解析している。落合は、Polysphondyliumの接着タンパク質抑制株を土台に、第2の接着タンパク質に対する特異抗体を調製し、新規接着タンパク質を同定しようとしている。細胞分化の分子機構前田(靖)は増殖・分化の切り替えの分子機構を知るために、細胞周期上の増殖分化の分岐点(PS点)前後で特異的に発現する遺伝子(Quit2,Quit3)について調べ、Quit2は新規のカルシュウム結合性タンパク質をコードしており、Quit3はアネキシンVII遺伝子の相補鎖をコードしていることを明らかにした。大畠は予定胞子細胞の分化誘導因子を単一標品に精製した。前田(ミ)は挿入突然変異法により得たMAPカイネースの欠損株を解析して、この遺伝子は集合に必須であること、またその活性調節には3量体Gタンパク質は関与していないことを明らかにした。田仲はPolysphondyliumの挿入突然変異体の作製方法を確立した。この方法を用いて得られた形質転換体を解析して、ベクターDNAがゲノムDNAにランダムに1コピーで挿入されているので有用な方法であると結論した。石田は単細胞で胞子に分化するsporogenous変異体と″固い集合体″で発生の停止するtag変異体を挿入突然変異体法で分離し、それらの遺伝子を同定した。小山は浸透圧ストレス耐性の変異株を挿入突然変異体により作製する条件を検討している。

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] Yumura,S.: "Intracellular free calcium responses during chemotaxis of Dictyostelium cells" J.Cell Sci.109. 2673-2678 (1996)
[文献書誌] Saito,T.: "Identification of a novel all-cis-5,9,12 heptadecatrienoic acid in the cellular slime mold Polysphondylium pallidum" Lipids,. 31. 445-447 (1996)
[文献書誌] Funamoto,S.: "Antisence RNA inactivation of gene expression of a cell-cell adhesion protein (gp64) in the cellular slime mold Polysphondylium pallidum" J.Cell Sci.109. 1009-1016 (1996)
[文献書誌] Fukuzawa,M.: "Molecular cloning and characterization of the cDNA for Discoidin II of Dictyostelium discoideum" Plant Cell Physiol.37. 505-514 (1996)
[文献書誌] Ochiai,H.: "Dimer formation of a cell-cell adhesion protein,gp64 of the cellular slime mold,Polysphondylium pallidum" Plant Cell Physiol. 37. 135-139 (1996)
[文献書誌] Nakata,N.: "Inhibitory Effects of Tunicamycin on Cell Cell Adhesion and Cell Reaggregation of the Cellular Slime Mold,Polysphondylium pallidum." J.Plant Res.109. 193-199 (1996)
[文献書誌] Amagai,A.: "Dictyostelium mucoroides cells exhibit cell cycle-dependent sorting behavior as observed in D.discoideum development" Dev.Growth & Differ.38. 33-40 (1996)
[文献書誌] Maeda,M.: "Seven helix chemoattractant receptors transiently stimulate mitogenactivated protein kinase in Dictyostelium" J.Biol.Chem.271. 3351-3354 (1996)
[文献書誌] Yamaguchi,N.,: "Targeted disruption of genes for gp138,a cell-fusion-related protein in Dictyostelium discoideum,revealed the existence of a third gene." Dev.Growth & Differ.38. 271-279 (1996)
[文献書誌] Pi,M.: "Analysis of a tRNA gene-like sequence (t-element) with TTA at the anticodon position in the mitochondrial DNA of Dictyostelium discoideum." J.Plant Res.109. 1-6 (1996)
[文献書誌] Urushihara,H.: "Protease-sensitive component (s) on cell surface prevents self-fusion in a bisexual strain of Dictyostelium discoideum." Cell Struct.Funct.21. 41-46 (1996)
[文献書誌] Urushihara,H.: "Choice of partners : Sexual cell interactions in Dictyostelium discoideum" Cell Struct.Funct.21. 231-236 (1996)
[文献書誌] Kuwayama,H.: "Aberrant cGMP-binding activity in non-chemotactic Dictyostelium discoideum mutants." Biochim.Biphys.Acta,. 1268. 214-217 (1996)
[文献書誌] Oyama,M.: "cGMP accumulation induced by hypertonic stress in Dictyostelium discoideum." J.Biol.Chem.271. 5574-5579 (1996)
[文献書誌] Yumura,S.: "Rapid redistribution of myosin II in living Dictyostelium amoebae,as revealed by fluorescent probes introduced by electroporation." Protoplasma. 192. 217-227 (1996)
[文献書誌] Yumura,S.: "Spatial distribution of fluorescently labeled actin in living Dictyostelium amoebae" Cell Struct. Funct.21. 189-197 (1996)
[文献書誌] Yumura,S.: "Myosin II can be localized to the cleavage furrow and to the posterior region of Dictyostelium amoebae without control by phosphorylation of myosin heavy and light chains." Cell Motil.Cytoskel.,in press. (1977)
[文献書誌] Fujita,H.: "Characterization of mutant myosins equivalent to human familial hypertophic cardiomyopathy mutants." J.Clinc.Invest.,in press. (1997)
[文献書誌] Arakane,T.: "Relevance of histone H1 kinase activity to the G2/M transition during the cell cycle of Dictyostelium discoideum." J.Plant Res.,in press. (1997)
[文献書誌] Okafuji,T.: "Antisense-mediated regulation of Annexin VII gene expression during the transition from growth to differentiation in Dictyostelium discoideum." Gene,in press. (1997)
[文献書誌] Aubry,L.: "The Dictyostelium MAP kinase ERK2 is regulated by Ras and cAMP-dependent protein kinase (PKA) and mediates PKA function." J.Biol.Chem.in press. (1997)
[文献書誌] Ogawa,S.: "A site-specific DNA endonuclease specified by one of two ORFs encoded by a group I intron in Dictyostelium discoideum mitochondrial DNA." Gene,in press. (1997)
[文献書誌] Komori,K.: "Localization of a DNA topoisomerase II to mitochondria in Dictyostelium discoideum : deletion mutant analysis and mitochondrial targeting signal presequence." J.Plant Res.,. (1997)
[文献書誌] Komori,K.: "Cloning and characterization of the gene encoding a mitochondrially localized DNA topoisomerase II in Dictyostelium discoideum : Western blot analysis." Biochim.Biophys.Acta,in press. (1997)
[文献書誌] Aiba,K.: "Isoforms of gp138,a cell-fusion related protein in Dictyostelium discoideum." J.Biochem.,in press. (1997)
[文献書誌] Ogawa,S.: "Group I introns in the cytochrome oxidase genes of Dictyostelium discoideum : two related ORFsin one loop of a group I intron,a cox1/2 hybrid gene and an unusually large cox3 gene." Curr.Genet.,in press. (1997)
[文献書誌] Shimizu,H.: "A mutation in the cAMP signaling pathway affects sexual development of Dictyostelium discoideum." Dev.Growth & Differ.,in press. (1997)
[文献書誌] 雨貝愛子: "「図解生物科学講座3:発生生物学」(浅島誠編著)" 朝倉書店,東京, 250 (1996)

1996 年度 実績報告書

挿入突然変異体による粘菌細胞の分化と形態形成の解析

研究代表者

竹内 郁夫 岡崎国立共同研究機構, 機構長 (90025239)

研究成果

[文献書誌] Yumura,S.: "Intracellular free calcium responses during chemotaxis of Dictyostelium cells" J.Cell Sci.109. 2673-2678 (1996)

[文献書誌] Saito,T.: "Identification of a novel all-cis-5,9,12 heptadecatrienoic acid in the cellular slime mold Polysphondylium pallidum" Lipids,. 31. 445-447 (1996)

[文献書誌] Funamoto,S.: "Antisence RNA inactivation of gene expression of a cell-cell adhesion protein (gp64) in the cellular slime mold Polysphondylium pallidum" J.Cell Sci.109. 1009-1016 (1996)

[文献書誌] Fukuzawa,M.: "Molecular cloning and characterization of the cDNA for Discoidin II of Dictyostelium discoideum" Plant Cell Physiol.37. 505-514 (1996)

[文献書誌] Ochiai,H.: "Dimer formation of a cell-cell adhesion protein,gp64 of the cellular slime mold,Polysphondylium pallidum" Plant Cell Physiol. 37. 135-139 (1996)

[文献書誌] Nakata,N.: "Inhibitory Effects of Tunicamycin on Cell Cell Adhesion and Cell Reaggregation of the Cellular Slime Mold,Polysphondylium pallidum." J.Plant Res.109. 193-199 (1996)

[文献書誌] Amagai,A.: "Dictyostelium mucoroides cells exhibit cell cycle-dependent sorting behavior as observed in D.discoideum development" Dev.Growth & Differ.38. 33-40 (1996)

[文献書誌] Maeda,M.: "Seven helix chemoattractant receptors transiently stimulate mitogenactivated protein kinase in Dictyostelium" J.Biol.Chem.271. 3351-3354 (1996)

[文献書誌] Yamaguchi,N.,: "Targeted disruption of genes for gp138,a cell-fusion-related protein in Dictyostelium discoideum,revealed the existence of a third gene." Dev.Growth & Differ.38. 271-279 (1996)

[文献書誌] Pi,M.: "Analysis of a tRNA gene-like sequence (t-element) with TTA at the anticodon position in the mitochondrial DNA of Dictyostelium discoideum." J.Plant Res.109. 1-6 (1996)

[文献書誌] Urushihara,H.: "Protease-sensitive component (s) on cell surface prevents self-fusion in a bisexual strain of Dictyostelium discoideum." Cell Struct.Funct.21. 41-46 (1996)

[文献書誌] Urushihara,H.: "Choice of partners : Sexual cell interactions in Dictyostelium discoideum" Cell Struct.Funct.21. 231-236 (1996)

[文献書誌] Kuwayama,H.: "Aberrant cGMP-binding activity in non-chemotactic Dictyostelium discoideum mutants." Biochim.Biphys.Acta,. 1268. 214-217 (1996)

[文献書誌] Oyama,M.: "cGMP accumulation induced by hypertonic stress in Dictyostelium discoideum." J.Biol.Chem.271. 5574-5579 (1996)

[文献書誌] Yumura,S.: "Rapid redistribution of myosin II in living Dictyostelium amoebae,as revealed by fluorescent probes introduced by electroporation." Protoplasma. 192. 217-227 (1996)

[文献書誌] Yumura,S.: "Spatial distribution of fluorescently labeled actin in living Dictyostelium amoebae" Cell Struct. Funct.21. 189-197 (1996)

[文献書誌] Yumura,S.: "Myosin II can be localized to the cleavage furrow and to the posterior region of Dictyostelium amoebae without control by phosphorylation of myosin heavy and light chains." Cell Motil.Cytoskel.,in press. (1977)

[文献書誌] Fujita,H.: "Characterization of mutant myosins equivalent to human familial hypertophic cardiomyopathy mutants." J.Clinc.Invest.,in press. (1997)

[文献書誌] Arakane,T.: "Relevance of histone H1 kinase activity to the G2/M transition during the cell cycle of Dictyostelium discoideum." J.Plant Res.,in press. (1997)

[文献書誌] Okafuji,T.: "Antisense-mediated regulation of Annexin VII gene expression during the transition from growth to differentiation in Dictyostelium discoideum." Gene,in press. (1997)

[文献書誌] Aubry,L.: "The Dictyostelium MAP kinase ERK2 is regulated by Ras and cAMP-dependent protein kinase (PKA) and mediates PKA function." J.Biol.Chem.in press. (1997)

[文献書誌] Ogawa,S.: "A site-specific DNA endonuclease specified by one of two ORFs encoded by a group I intron in Dictyostelium discoideum mitochondrial DNA." Gene,in press. (1997)

[文献書誌] Komori,K.: "Localization of a DNA topoisomerase II to mitochondria in Dictyostelium discoideum : deletion mutant analysis and mitochondrial targeting signal presequence." J.Plant Res.,. (1997)

[文献書誌] Komori,K.: "Cloning and characterization of the gene encoding a mitochondrially localized DNA topoisomerase II in Dictyostelium discoideum : Western blot analysis." Biochim.Biophys.Acta,in press. (1997)

[文献書誌] Aiba,K.: "Isoforms of gp138,a cell-fusion related protein in Dictyostelium discoideum." J.Biochem.,in press. (1997)

[文献書誌] Ogawa,S.: "Group I introns in the cytochrome oxidase genes of Dictyostelium discoideum : two related ORFsin one loop of a group I intron,a cox1/2 hybrid gene and an unusually large cox3 gene." Curr.Genet.,in press. (1997)

[文献書誌] Shimizu,H.: "A mutation in the cAMP signaling pathway affects sexual development of Dictyostelium discoideum." Dev.Growth & Differ.,in press. (1997)

[文献書誌] 雨貝愛子: "「図解生物科学講座3:発生生物学」(浅島誠 編著)" 朝倉書店,東京, 250 (1996)

1996 年度実績報告書

竹内郁夫岡崎国立共同研究機構, 機構長 (90025239)

[文献書誌] 雨貝愛子: "「図解生物科学講座3:発生生物学」(浅島誠編著)" 朝倉書店,東京, 250 (1996)