1997 Fiscal Year Annual Research Report

LIMキナーゼの分子細胞生物学的研究ならびに遺伝子欠損マウス作製による機能解析

Research Project

Project/Area Number	09670127
Research Category	Grant-in-Aid for Scientific Research (C)
Research Institution	Osaka University
Principal Investigator	松本邦夫大阪大学, 医学部, 助教授 (90201780)
Co-Investigator(Kenkyū-buntansha)	船越洋大阪大学, 医学部, 助手 (40273685) 宮本一政大阪大学, 医学部, 助手 (60273679) 小清水右一大阪大学, 医学部, 助手 (50281126)
Keywords	LIMキナーゼ / yeast two-hybrid / POZドメイン / ウィリアム症候群 / プロテインキナーゼ
Research Abstract	LIMキナーゼ(LINK)は我々のグループが同定した新規のセリン/チロシンキナーゼであり、LIMK1とLIMK2の2つのサブタイプが存在する。LINKは2つのLIMドメインとPDZドメインを併せもつなど他に類を見ない構造的特徴を有する。成体においては、LIMK1は脳・神経組織に限局した発現が認められる一方、LIMK2は脳を含む広い組織で発現されている。最近LIMK1は発現認知能に異常をきたす遺伝子疾患ウイリアムス症候群の原因遺伝子であることが明らかにされた。しかしながら、その発症機構を含めてLIMKの機能はほとんど不明であり、その機能を明らかにする上で基質や活性化因子の同定は不可欠なものである。今回、Yeast Two-Hybrid法を用いて既知・新規蛋白質をあわせて56種ものLIMK相互作用分子を同定した。LIMK1の相互作用分子として既知分子を18クローン、新規分子を9クローン同定したが、既知分子の中には、アクチンに結合する細胞骨格系蛋白質、DNA複製装置を構成する蛋白質、ならびに新規のZinc-Finger蛋白を含む数種の転写因子が含まれている。一方、LIMK2からは既知分子を14クローン、新規分子を15クローン同定した。その中には、やはり細胞骨格系の蛋白質や転写因子が含まれる他、核分裂装置を構成する蛋白質やWntカスケードに関わる分子などが含まれている。マウスLIMK2は様々な組織で発現しているが、LIMドメインを欠失した精巣特異的LIMK2バリアントは減数分裂後精母細胞や精子細胞に限局した発現がみられた。 Yeast Two-Hybrid法によってLIMKと相互作用する分子が予想をはるかに上回る種類同定された。これまでにLIMドメインをもつ蛋白質は細胞骨格の制御に関与するもの、核内転写因子として機能するものが知られていることから、LIM蛋白質群は細胞形態の変化によって引き起こされる増殖・分化の制御や遺伝子発現の切り換えといった未知の情報伝達系に関与することが推定された。実際にLIMK相互作用分子の中には細胞骨格系蛋白質や転写因子が多く含まれており、正常細胞とがん細胞を区別するanchorage dependencyを制御する情報伝達、細胞形態変化の察知から遺伝子発現の切り換えに至る情報伝達系などにおいて、LIMKは重要な機能を果たすことが予想される。また、LIMK1にはDNA複製装置、LIMK2には核分裂装置の蛋白質が相互作用しており、LIMKが細胞周期の制御にも密接に関与している可能性が示された。 LIMK2はそのファミリー分子であるLIMK1との同様、脳・神経発生における重要性が示唆されているもののその機能はほとんどわかっていない。今回、精巣特異的に発現するLIMK2バリアント、tLIMK2を同定し、その発現が減数分裂期の生殖細胞に限局していることを明らかにしたが、tLIMK2は細胞内局在や活性化状態が通常のLIMK2分子とは異なることが示唆され、生殖細胞形成、特に減数分裂制御過程において重要な役割を担うものと考えられる。

Research Products

(30 results)

All Other

All Publications (30 results)

[Publications] K.Takai: "Hepatocyte growth factor is constitutively produced by human bone marrow stromal cells and promotes erythropoiesis." Blood. 89. 1560-1565 (1997)
[Publications] R.Morishita: "Potential role of endothelium-specific growth factor hepatocyte growth factor,on endothelial damage in diabetes militus." Diabetes. 46. 138-142 (1997)
[Publications] N.Kawakami-Kimura: "Involvement of hepatocyte growth factor in increased integrin expression on Hep G2 cells triggered by adhesion to endothelial cells." Br.J.Cancer. 75. 47-53 (1997)
[Publications] T.Takahashi: "Xenopus LIM motif-containing protein kinase,XLIMK,is expressed in the developing head structure of the embryo." Develop.Dyn.209. 196-205 (1997)
[Publications] S.Aoki: "Activation of Met tyrosine kinase by hepatocyte growth factor is essential for internal ganogenesis in Xenopus embryo." Biochem.Biophys.Res.Commun.234. 8-14 (1997)
[Publications] Y.Matsuda: "Prevential and therapeutic effects in rats of hepatocyte growth factor infusion on liver fibrosis/cirrhosis." Hepatology. 26. 81-89 (1997)
[Publications] T.Nakamura: "Induction of hepatocyte growth factor in fibroblasts by tumor-derived factors affects invasive growth of tumor cells : In vitro analysis of tumor-stromal interaction." Cancer Res.57. 3305-3313 (1997)
[Publications] R.Morishita: "Role of hepatocyte growth factor in endothelial regulatoin : Prevention of high D-glucose induced endothelial cell death by prostglandins and phosphodiesterase type 3 inhibitor." Diabetologia. 40. 1053-1061 (1997)
[Publications] H.Li: "Hepatocyte gorwht factor stimuolates invasion of gallbladder carcinoma cell lines in vitro." Clin.Exp.Metastasis.15. 484-493 (1997)
[Publications] S.Wakitani: "Hepatocyte growth factor faulitates Cartilage repair : full thickness articular cartilage defect studied in rabbit knee." Acta Orthop.Scand.68. 474-480 (1997)
[Publications] N.Nakano: "Role of antagotensin II in the reguration of a novel vascular modulator,hepatocyte growth factor (HGF),in experimental hypertensive rats." Hypertension. 30. 1448-1454 (1997)
[Publications] K.Date: "HGF/NK4 is a specific antagonist for pleiotrophic actions of hepatocyte growth factor." FEBS Lett.420. 1-6 (1997)
[Publications] A.Schmassman: "Roles of hepatocyte growth facotr and its receptor c-met during gastric ulcer healing in rats." Gastroenterolgy. 113. 1858-1872 (1997)
[Publications] M.Yaekashiwa: "Both simultaneous and delayed administration of hepatocyte growth factor (HGF) repress the fibrotic changes in murine lung injury induced by bleomycin." Am.J.Resp.& Clit.Care Med.156. 1937-1944 (1997)
[Publications] Y.Yo: "Actions of hepatocyte growth factor as a local modulator in the kidney : Potential role in pathogenesis of renal disease." Kidney Intern.53. 50-58 (1998)
[Publications] K.Matsumoto: "Hepatocyte growth factor (HGF) as a tissue organizer for organogenesis and regeneration." Biochem.Biophys.Res.Commun.239. 639-644 (1997)
[Publications] R.Morishita: "Potential role of a novel vascular modulator,hepatocyte growth factor (HGF),in cardiovascular disease : characterization and regulation of local HGF system." J.Atheroscle.Thromb.4. 12-19 (1997)
[Publications] S.Nakamura: "A novel vascular modulator,hepatocyte growth factor (HGF),as a potential index of the severity of hypertension." Biochem.Biophys.Res.Commun.242. 238-243 (1998)
[Publications] K.Liu: "Protamine enhances the proliferative activity of hepatocyte growth factor in rats." Am.J.Physiol.(in press). (1998)
[Publications] T.Miyazawa: "Protection of hippocampal neurons from ischemia-induced delayed neuronal death by hepatocyte growth factor : A novel neurotrophic factor." J.Celebral Blood Flow Metabolism.(in press). (1998)
[Publications] H.Ohmichi: "Hepatocyte growth factor (HGF) acts as a mesenchynal-derived morphogenic factor during fetal lung development." Development.(in press). (1998)
[Publications] S.Mizuno: "Hepatocyte growth factor prevents renal fibrosis and dysfunction in mouse model of chronio renal disease." J.Clin.Invest.(in press). (1998)
[Publications] K.Kosai: "Hepatocyte growth factor abrogates Fas-induced lethal livel poptosis in mice." Biochem.Biophys.Res.Commun.(in press). (1998)
[Publications] K.Matsumoto and T.Nakamura: "Jhon Wiley & Sons" Plasminogen-Related Growth Factors, 198-214 (1997)
[Publications] K.Matsumoto and T.Nakamura: "Internal Medicine and Endocrinology" Pathophysiological and Clinical Interactions, 95-106 (1997)
[Publications] 松本邦夫、中村敏一: "癌・間質相互作用(tumor-stromal interaction)" 文光堂, 40 (1997)
[Publications] 松村邦夫、中村敏一: "HGFの臨床応用:難治性疾患療薬の新しいターゲット" 羊土社, 90-97 (1997)
[Publications] 大道栄徳、松本邦夫: "メディカルレビュー社" 肺とHGF : 肺再生機構から肺疾患治療へ, 51-56 (1997)
[Publications] 松本邦夫、中村敏一: "肝臓の発生と分化" 現代医療社, 79-86 (1997)
[Publications] 松本邦夫、中村敏一: "タンパク質の生化学的解析" 南江堂(印刷中), (1998)

1997 Fiscal Year Annual Research Report

LIMキナーゼの分子細胞生物学的研究ならびに遺伝子欠損マウス作製による機能解析

Principal Investigator

松本 邦夫 大阪大学, 医学部, 助教授 (90201780)

Research Products

[Publications] K.Takai: "Hepatocyte growth factor is constitutively produced by human bone marrow stromal cells and promotes erythropoiesis." Blood. 89. 1560-1565 (1997)

[Publications] R.Morishita: "Potential role of endothelium-specific growth factor hepatocyte growth factor,on endothelial damage in diabetes militus." Diabetes. 46. 138-142 (1997)

[Publications] N.Kawakami-Kimura: "Involvement of hepatocyte growth factor in increased integrin expression on Hep G2 cells triggered by adhesion to endothelial cells." Br.J.Cancer. 75. 47-53 (1997)

[Publications] T.Takahashi: "Xenopus LIM motif-containing protein kinase,XLIMK,is expressed in the developing head structure of the embryo." Develop.Dyn.209. 196-205 (1997)

[Publications] S.Aoki: "Activation of Met tyrosine kinase by hepatocyte growth factor is essential for internal ganogenesis in Xenopus embryo." Biochem.Biophys.Res.Commun.234. 8-14 (1997)

[Publications] Y.Matsuda: "Prevential and therapeutic effects in rats of hepatocyte growth factor infusion on liver fibrosis/cirrhosis." Hepatology. 26. 81-89 (1997)

[Publications] T.Nakamura: "Induction of hepatocyte growth factor in fibroblasts by tumor-derived factors affects invasive growth of tumor cells : In vitro analysis of tumor-stromal interaction." Cancer Res.57. 3305-3313 (1997)

[Publications] R.Morishita: "Role of hepatocyte growth factor in endothelial regulatoin : Prevention of high D-glucose induced endothelial cell death by prostglandins and phosphodiesterase type 3 inhibitor." Diabetologia. 40. 1053-1061 (1997)

[Publications] H.Li: "Hepatocyte gorwht factor stimuolates invasion of gallbladder carcinoma cell lines in vitro." Clin.Exp.Metastasis.15. 484-493 (1997)

[Publications] S.Wakitani: "Hepatocyte growth factor faulitates Cartilage repair : full thickness articular cartilage defect studied in rabbit knee." Acta Orthop.Scand.68. 474-480 (1997)

[Publications] N.Nakano: "Role of antagotensin II in the reguration of a novel vascular modulator,hepatocyte growth factor (HGF),in experimental hypertensive rats." Hypertension. 30. 1448-1454 (1997)

[Publications] K.Date: "HGF/NK4 is a specific antagonist for pleiotrophic actions of hepatocyte growth factor." FEBS Lett.420. 1-6 (1997)

[Publications] A.Schmassman: "Roles of hepatocyte growth facotr and its receptor c-met during gastric ulcer healing in rats." Gastroenterolgy. 113. 1858-1872 (1997)

[Publications] M.Yaekashiwa: "Both simultaneous and delayed administration of hepatocyte growth factor (HGF) repress the fibrotic changes in murine lung injury induced by bleomycin." Am.J.Resp.& Clit.Care Med.156. 1937-1944 (1997)

[Publications] Y.Yo: "Actions of hepatocyte growth factor as a local modulator in the kidney : Potential role in pathogenesis of renal disease." Kidney Intern.53. 50-58 (1998)

[Publications] K.Matsumoto: "Hepatocyte growth factor (HGF) as a tissue organizer for organogenesis and regeneration." Biochem.Biophys.Res.Commun.239. 639-644 (1997)

[Publications] R.Morishita: "Potential role of a novel vascular modulator,hepatocyte growth factor (HGF),in cardiovascular disease : characterization and regulation of local HGF system." J.Atheroscle.Thromb.4. 12-19 (1997)

[Publications] S.Nakamura: "A novel vascular modulator,hepatocyte growth factor (HGF),as a potential index of the severity of hypertension." Biochem.Biophys.Res.Commun.242. 238-243 (1998)

[Publications] K.Liu: "Protamine enhances the proliferative activity of hepatocyte growth factor in rats." Am.J.Physiol.(in press). (1998)

[Publications] T.Miyazawa: "Protection of hippocampal neurons from ischemia-induced delayed neuronal death by hepatocyte growth factor : A novel neurotrophic factor." J.Celebral Blood Flow Metabolism.(in press). (1998)

[Publications] H.Ohmichi: "Hepatocyte growth factor (HGF) acts as a mesenchynal-derived morphogenic factor during fetal lung development." Development.(in press). (1998)

[Publications] S.Mizuno: "Hepatocyte growth factor prevents renal fibrosis and dysfunction in mouse model of chronio renal disease." J.Clin.Invest.(in press). (1998)

[Publications] K.Kosai: "Hepatocyte growth factor abrogates Fas-induced lethal livel poptosis in mice." Biochem.Biophys.Res.Commun.(in press). (1998)

[Publications] K.Matsumoto and T.Nakamura: "Jhon Wiley & Sons" Plasminogen-Related Growth Factors, 198-214 (1997)

[Publications] K.Matsumoto and T.Nakamura: "Internal Medicine and Endocrinology" Pathophysiological and Clinical Interactions, 95-106 (1997)

[Publications] 松本邦夫、中村敏一: "癌・間質相互作用(tumor-stromal interaction)" 文光堂, 40 (1997)

[Publications] 松村邦夫、中村敏一: "HGFの臨床応用:難治性疾患療薬の新しいターゲット" 羊土社, 90-97 (1997)

[Publications] 大道栄徳、松本邦夫: "メディカルレビュー社" 肺とHGF : 肺再生機構から肺疾患治療へ, 51-56 (1997)

[Publications] 松本邦夫、中村敏一: "肝臓の発生と分化" 現代 医療社, 79-86 (1997)

[Publications] 松本邦夫、中村敏一: "タンパク質の生化学的解析" 南江堂(印刷中), (1998)

松本邦夫大阪大学, 医学部, 助教授 (90201780)

[Publications] 松本邦夫、中村敏一: "肝臓の発生と分化" 現代医療社, 79-86 (1997)