2001 年度実績報告書

細胞質分裂期におけるリン酸化ミオシンIIの役割解明に関する研究

研究課題

研究課題/領域番号	10213206
研究機関	広島大学
研究代表者	細谷浩史広島大学, 大学院・理学研究科, 教授 (90183102)
研究分担者	祐村恵彦山口大学, 理学部, 助教授 (70183986)
キーワード	ミオシン調節軽鎖 / ミオシン重鎖 / 細胞骨格 / アクチン / ミオシンII / リン酸化 / ストレスファイバー / 収縮環
研究概要	細胞分裂期におけるミオシンリン酸化の役割解明を目的として、ヒーラ細胞を実験材料に使用して以下の成果を得た。 1.19Serがリン酸化されたミオシン調節軽鎖を特異的に認識するモノクローナル抗体を作製した。 2.18Thrおよび19Serの2ケ所が2重リン酸化されたミオシン調節軽鎖を特異的に認識するモノクローナル抗体を作成した。 3.これらの抗体を用いて、間期および分裂期のヒーラ細胞を染色したところ、間期の細胞ではストレスファイバーが染色され、分裂期の細胞では収縮環の領域が染色されることが明らかになった。 4.2重リン酸化ミオシンに対する抗体およびミオシン重鎖の抗体を用いて分裂期の細胞を2重染色したところ、収縮環領域では、前者はdisc状に染色像が観察されたが、後者の染色像はリング状であることが明らかになった。これらの結果から、収縮環でミオシン重鎖と軽鎖が必ずしも共局在しないという事実が明らかにされた。 5.ミオシン調節軽鎖のリン酸化部位に関する変異体を作成しヒーラ細胞への導入を行ったところ、18Thrおよび19Serにアスパラギン酸を導入した疑似2重リン酸化ミオシン調節軽鎖を導入した細胞では、細胞分裂の阻害は特に観察されないが、これらの変異ミオシン調節軽鎖は収縮環領域に局在することが明らかにされた。19Serにアスパラギン酸を導入した疑似1重リン酸化ミオシン調節軽鎖の場合でも同様の結果が観察されたが、収縮環への局在は前者ほど顕著ではなかった。また野生型ミオシン調節軽鎖を導入した場合では、収縮環への局在は顕著に観察されなかった。

研究成果
(24件)

すべてその他

すべて文献書誌 (24件)

[文献書誌] M.A.Okubo: "A monoclonal antibody, mHl, specifically recognizes micrerilli of dividing HeLa cells"Tiss.Cult.Res.Commun.. 18. 113-117 (1999)
[文献書誌] G.Totsukawa: "Activation of myosin phosphatase targeting subunit by mitosis-specific phosphorylation"J.Cell Biol.. 144(4). 735-744 (1999)
[文献書誌] M.A.Okubo: "Generation and characterization of a monoclonal antibody, mHl, raised against mitotic HeLa cells"Dev.Growth Differ.. 41(4). 381-389 (1999)
[文献書誌] N.Nishihara: "Microtubule-dependent movement of symbiotic algae and granules in Paramecium bursaria"Cell Motil.Cytoskeleton.. 43(2). 85-98 (1999)
[文献書誌] M.Murata-Hori: "ZIP kinase identified as a novel myosin regulatory light chain kinase in HeLa cells"FEBS Lett.. 451(1). 81-84 (1999)
[文献書誌] M.Murata-Hori: "A monoclonal antibody raised against an 86-kD subunit of human DNA helicase II"Zool.Sci.. 17(9). 1255-1259 (2000)
[文献書誌] M.Murata-Hori: "Myosin II regulatory light chain as a novel substrate for AIM-l, an Aurora/Ipllp-related kinase from rat"J.Biochem.. 128(6). 903-907 (2000)
[文献書誌] B.I.Gerashchenko: "Flow cytometry as a strategy to study the endosymbiosis of algae in Paramecium bursaria"Cytometry. 41(3). 209-215 (2000)
[文献書誌] B.I.Gerashchenko: "Enrichment for late-telophase cell populations using flow cytometry"Cytometry. 41(2). 148-149 (2000)
[文献書誌] F.Suizu: "Activation of actin-activated MgATPase activity of myosin II by phosphorylation with MAPK-activated protein kinase-1b (RSK-2)"J.Biochem.. 128(3). 436-440 (2000)
[文献書誌] B.I.Gerashchenko: "Myosin regulatory light chain as a critical substrate of cell death : a hypothesis"Med.Hypoth.. 54(5). 850-852 (2000)
[文献書誌] M.Hino: "Small heat shock protein 27(HSP27) associates with tubulin/microtubules in HeLa cells"Biochem.Biophys.Res.Commun.. 271(1). 146-169 (2000)
[文献書誌] T.Saitoh: "Differential localization of non-muscle myosin II isoforms and phosphorylated regulatory light chains in human MIRC-5 fibroblasts"FEBS Lett.. 509. 365-369 (2001)
[文献書誌] M.Murata-Hori: "HeLaZIP kinase induces diphosphorylation of Myosin II regulatory light chain and reorganization of actin filaments in nonmuscle cells"Oncogene. 20. 8175-8183 (2001)
[文献書誌] T.Kawano: "A Fungal anxin antagonist, hypaphorine competitively inhibits indole-3-acetic acid-dependent superoxide generation by horseradish peroxidase"Biochem.Biophys.Res.Commun.. 288. 546-551 (2001)
[文献書誌] B.I.Gerashchenko: "Model for regulation of non-muscle myosnis"Fiziologichny Zhrbal. 47(2). 100-105 (2001)
[文献書誌] S.Komaba: "Identificafion of myosin II kinase from sea urchin eggs as protein kinase CK2"Gene. 275(1). 141-148 (2001)
[文献書誌] B.I.Gerashchenko: "Growth kinetics of algal populations exsymbiotic from Paramecium bursaria by flow cytometry measurements"Cytometry. 44(3). 257-263 (2001)
[文献書誌] B.I.Gerashchenko: "On regulation of nonmuscle myosin-II"Recent Research Developments in Biophysics and Biochemistry. 1. 1-7 (2001)
[文献書誌] B.I.Gerashchenko: "New horizons for endosymbiosis of algae in Paramecium bursaria in light of flow"Recent Research Developments in Biophysics and Biochemistry. 1. 145-160 (2001)
[文献書誌] T.Kawano: "Spectroscopic evidence that salicylic acid converts a temporal inactive form of horseradish peroxidase(Compound III) to the irreversibly inactivated verdohemoprotein(P-670)"Biosci Biotechnol.Biochem.. 66(3). 646-650 (2002)
[文献書誌] B.I.Gerashchenko: "Phosphorylation at threonine-18 in addition to phosphorylation at Serine-19 on myosin-II regulatory light chain is a mitosis-specific event"Cytometry. 47. 150-157 (2002)
[文献書誌] T.Iwasaki: "Diphosphorylated MRLC is required for organization of stress fibers in interphase cells and the contractile ring in dividing cells"Cell struc.Funct.. 26. 677-683 (2001)
[文献書誌] T.Kawano: "Spectroscopic evidence in support of horseradish peroxidase Compound II-catalyzed oxidation of salicylic acid but not of phenylethylamine"Biosci.Biotechnol.Biochem.. 66(3). 651-654 (2002)

2001 年度 実績報告書

細胞質分裂期におけるリン酸化ミオシンIIの役割解明に関する研究

研究代表者

細谷 浩史 広島大学, 大学院・理学研究科, 教授 (90183102)

研究成果

[文献書誌] M.A.Okubo: "A monoclonal antibody, mHl, specifically recognizes micrerilli of dividing HeLa cells"Tiss.Cult.Res.Commun.. 18. 113-117 (1999)

[文献書誌] G.Totsukawa: "Activation of myosin phosphatase targeting subunit by mitosis-specific phosphorylation"J.Cell Biol.. 144(4). 735-744 (1999)

[文献書誌] M.A.Okubo: "Generation and characterization of a monoclonal antibody, mHl, raised against mitotic HeLa cells"Dev.Growth Differ.. 41(4). 381-389 (1999)

[文献書誌] N.Nishihara: "Microtubule-dependent movement of symbiotic algae and granules in Paramecium bursaria"Cell Motil.Cytoskeleton.. 43(2). 85-98 (1999)

[文献書誌] M.Murata-Hori: "ZIP kinase identified as a novel myosin regulatory light chain kinase in HeLa cells"FEBS Lett.. 451(1). 81-84 (1999)

[文献書誌] M.Murata-Hori: "A monoclonal antibody raised against an 86-kD subunit of human DNA helicase II"Zool.Sci.. 17(9). 1255-1259 (2000)

[文献書誌] M.Murata-Hori: "Myosin II regulatory light chain as a novel substrate for AIM-l, an Aurora/Ipllp-related kinase from rat"J.Biochem.. 128(6). 903-907 (2000)

[文献書誌] B.I.Gerashchenko: "Flow cytometry as a strategy to study the endosymbiosis of algae in Paramecium bursaria"Cytometry. 41(3). 209-215 (2000)

[文献書誌] B.I.Gerashchenko: "Enrichment for late-telophase cell populations using flow cytometry"Cytometry. 41(2). 148-149 (2000)

[文献書誌] F.Suizu: "Activation of actin-activated MgATPase activity of myosin II by phosphorylation with MAPK-activated protein kinase-1b (RSK-2)"J.Biochem.. 128(3). 436-440 (2000)

[文献書誌] B.I.Gerashchenko: "Myosin regulatory light chain as a critical substrate of cell death : a hypothesis"Med.Hypoth.. 54(5). 850-852 (2000)

[文献書誌] M.Hino: "Small heat shock protein 27(HSP27) associates with tubulin/microtubules in HeLa cells"Biochem.Biophys.Res.Commun.. 271(1). 146-169 (2000)

[文献書誌] T.Saitoh: "Differential localization of non-muscle myosin II isoforms and phosphorylated regulatory light chains in human MIRC-5 fibroblasts"FEBS Lett.. 509. 365-369 (2001)

[文献書誌] M.Murata-Hori: "HeLaZIP kinase induces diphosphorylation of Myosin II regulatory light chain and reorganization of actin filaments in nonmuscle cells"Oncogene. 20. 8175-8183 (2001)

[文献書誌] T.Kawano: "A Fungal anxin antagonist, hypaphorine competitively inhibits indole-3-acetic acid-dependent superoxide generation by horseradish peroxidase"Biochem.Biophys.Res.Commun.. 288. 546-551 (2001)

[文献書誌] B.I.Gerashchenko: "Model for regulation of non-muscle myosnis"Fiziologichny Zhrbal. 47(2). 100-105 (2001)

[文献書誌] S.Komaba: "Identificafion of myosin II kinase from sea urchin eggs as protein kinase CK2"Gene. 275(1). 141-148 (2001)

[文献書誌] B.I.Gerashchenko: "Growth kinetics of algal populations exsymbiotic from Paramecium bursaria by flow cytometry measurements"Cytometry. 44(3). 257-263 (2001)

[文献書誌] B.I.Gerashchenko: "On regulation of nonmuscle myosin-II"Recent Research Developments in Biophysics and Biochemistry. 1. 1-7 (2001)

[文献書誌] B.I.Gerashchenko: "New horizons for endosymbiosis of algae in Paramecium bursaria in light of flow"Recent Research Developments in Biophysics and Biochemistry. 1. 145-160 (2001)

[文献書誌] T.Kawano: "Spectroscopic evidence that salicylic acid converts a temporal inactive form of horseradish peroxidase(Compound III) to the irreversibly inactivated verdohemoprotein(P-670)"Biosci Biotechnol.Biochem.. 66(3). 646-650 (2002)

[文献書誌] B.I.Gerashchenko: "Phosphorylation at threonine-18 in addition to phosphorylation at Serine-19 on myosin-II regulatory light chain is a mitosis-specific event"Cytometry. 47. 150-157 (2002)

[文献書誌] T.Iwasaki: "Diphosphorylated MRLC is required for organization of stress fibers in interphase cells and the contractile ring in dividing cells"Cell struc.Funct.. 26. 677-683 (2001)

[文献書誌] T.Kawano: "Spectroscopic evidence in support of horseradish peroxidase Compound II-catalyzed oxidation of salicylic acid but not of phenylethylamine"Biosci.Biotechnol.Biochem.. 66(3). 651-654 (2002)

2001 年度実績報告書

細谷浩史広島大学, 大学院・理学研究科, 教授 (90183102)