1995 年度実績報告書

抗-細胞接着の分子機構と生理機能の解析

研究課題

研究課題/領域番号	07308073
研究種目	総合研究(A)
研究機関	愛知医科大学
研究代表者	木全弘治愛知医科大学, 分子医科学研究所, 教授 (10022641)
研究分担者	祖父江憲治大阪大学, 医学部, 教授 (20112047) 関口清俊大阪府立母子保険総合医療センター, 部長 (50187845) 坂倉照よ三重大学, 医学部, 教授 (80073120) 北川泰雄生物分子応答研究センター, 教授 (50101168) 永田和宏京都大学, 胸部疾患研究所, 教授 (50127114)
キーワード	細胞接着 / 抗-接着 / インテグリン / 細胞基質相互作用 / プロテオグリカン / 接着シグナル / キナーゼ / レセプター
研究概要	研究代表者である木全は、コンドロイチン硫酸プロテオグリカンの抗-接着活性部位であるコンドロイチン硫酸鎖に対する細胞表面レセプターとしてannexineVIを同定した。また、コンドロイチン硫酸鎖による抗-細胞接着活性が、細胞内シグナル伝達系を介したものであることを3種類の解析法を考案して証明し、抗-細胞接着活性のレセプターとしてannexinVIの関与を調べる基礎を確立した。抗-接着活性を検出しているマトリックス分子として、プロテオグリカン(木全、大平、野田)、ラミニン(北川)、テネイシン(坂倉、小原)に注目した。これらの分子の抗-接着活性の発現には、多量体形成や分泌などの分子のプロセシングに依存する。その過程にシャペロンHSP47がコラーゲン分子の合成量の制御までも行うことを明らかにした(永田)。テネイシンの抗-接着活性は、二価金属イオンにより制御されることを明らかにした(小原)。抗-接着活性を示す分子の生理機能を、軟骨分化(木全)、癌転移(坂倉)、血管新生(北川)、神経回路網形成(大平、野田)、平滑筋分化(祖父江)において解析し、抗-細胞接着活性の重要性を示した。インテグリンやシンデカンの接着活性の制御機構の解析から(関口、岡山)、ラミニンによる細胞分化因子としての作用の解析から(祖父江)、ホスファターゼ活性を持つプロテオグリカンによるチロシンりん酸化レベルの制御機構の解析から(野田)、特異抗体による細胞接着に作用する新規のキナーゼの同定法を確立して中間径フィラメントのりん酸化反応の解析から(稲垣)、低分子GTP結合蛋白質Rhoについて2種のセリンスレオニンキナーゼと3種の非キナーゼを標的としたインテグリン活性化機構の解析から(成宮)、接着分子オクルディンのマウスcDNAの単離により遺伝子工学的解析を可能にして(月田)、インテグリンなどの接着分子の活性調整機構の存在や分子実体の解析に成功した。さらに、今年度の班会議を名古屋で開催し、班員相互の研究交流を行った。

研究成果

(47件)

すべてその他

すべて文献書誌 (47件)

[文献書誌] Ito, K. et al.: "Multiple forms of mouse PG-M, a large chondroitin sufate proteoglycan generated by alternative splicing" The Journal of Biological Chemistry. 270. 958-965 (1995)
[文献書誌] Zako, M. et al.: "Expression of PG-M(V3), an alternatively spliced form of PG-M without a chondroitin sulfate attachment region in mouse and human tissue." The Journal of Biological Chemistry. 270. 3914-3918 (1995)
[文献書誌] Shinomura, T. et al.: "The gene structure and organization for mouse PG-M, a large chondroitin sulfate proteoglycan: Genomic background for the generation of multiple PG-M transcripts" The Journal of Biological Chemistry. 270. 10328-10333 (1995)
[文献書誌] Kato, S. et. al.: "Chondroitin sulfate immobilized onto culture substrates modulates 6DNA synthesis, tyrosine aminotransferase induction, and intercellular communication in primary rat hepatocytes" Cell Structure and function. 20. 199-209 (1995)
[文献書誌] Fukuoka, M. et al.: "Molecular cloning and expression of chick chondrocyte chondroitin 6-sulfotransferase" The Journal of Biological Chemistry. 270. 18575-18580 (1995)
[文献書誌] Nagai, N. et al.: "Quercetin suppresses heat shock response by down regulation of HSP1" Biochemical and Biophysical Research Communications. 208. 1099-1105 (1995)
[文献書誌] Higashi, T. et al.: "Activation of heat shock factor 1 in rat brain during cerebral ischemia or after heat shock" Mol. Brain Res.34. 262-270 (1995)
[文献書誌] Nakai A. et al.: "The DNA-Binding properties of two heat shock factors, HSF1 and HSF3, are induced in the avian erythroblast cell line HD6" Molecular and Cellular Biology. 15. 5268-5278 (1995)
[文献書誌] Yasuda, K. et al.: "Cloning and expression of murine high-molecular-weight heat shock proteins, HSP105" The Journal of Biological Chemistry. 270. 29718-29723 (1995)
[文献書誌] Nagata, K. et al.: "HSP47: a collagen-specific molecular chaperone in the endoplasmic reticulum" Trends in Biochemical Sciences. 21. 23-26 (1996)
[文献書誌] Satoh, M. et al.: "Intracellular interaction of collagen-specific stress protein HSP47 with newly synthesized procollagen" The Journal of Cell Biology. (in press). (1996)
[文献書誌] Jeon, H. et al.: "Pericytes from microvessel fragment produce type IV collagen and multiple laminin isoforms" Biosci. Biotech. Biochem.(in press). (1995)
[文献書誌] Kumagai, C. et al.: "Laminin α3-antiserum recognizes α chain in laminin 5 (kalinnin) but not that in laminin 6 (K-laminin) of human keratinocytes" Biochim. Biophys. Acta. (in press). (1995)
[文献書誌] Watanabe, E. et al.: "Distribution of brain-specific proteoglycan, neurocan, and the correspondion mRNA during the formation of barrels in the rat" European Journal of Neuroscience. 7. 547-554 (1995)
[文献書誌] Katoh-Semba, R. et al.: "Chondroitin sulfate proteoglycan in the rat brain: Candidates for axon barriers of sensory neurons and the possible" Nerosci. Let.189. 25-28 (1995)
[文献書誌] Maeda, N. et al.: "Purfication, characterization and developmental expression of brain-specific chondroitin sulfate-" Neurosci.67. 23-35 (1995)
[文献書誌] Watanabe, E. et al.: "Neuroglycan C, a novel membrane-spanning chondroitin sulfate proteoglycan that is restricted to the brain" Journal of Biological Chemistry. 270. 26876-26882 (1995)
[文献書誌] 大平敦彦: "神経回路網形成期の脳に発現しているプロテオグリカンの構造と機能" 日本神経精神薬理学雑誌. 171-176 (1995)
[文献書誌] Mizutani, H. et al.: "Nodular scleroderma: Focally increased tenascin expession differing from that in the surrounding scleroderma soin" J. Dermatology. 22. 267-271 (1995)
[文献書誌] Ishihara, A. et al.: "Tenascin expressuin in cancer cells and stroma of human breast cancer amd its prognostic significance" Clin. Cancer Res.1. 1035-1041 (1995)
[文献書誌] Yoshida, T. et al.: "Tenascin expression in breast cancer cell: A maker for poor grognosis in breast cancer" Cancer Letter. 90. 65-73 (1995)
[文献書誌] Ichihara-Tanaka, K. et al.: "Role of the carboxyl-terminal Fib2 domain in fibronectin matrix assembly" Journal of Cell Science. 108. 907-915 (1995)
[文献書誌] Healy J. M. et al.: "Peptide ligands for integrin αvβ3 selected from random phage display libraries" Biochemistry. 34. 3948-3955 (1995)
[文献書誌] Ohashi, H. et al.: "Up-regulation of integrin α5β1 expression by interleukin-6 in rabbit corneal epithelial cells" Experimental Cell Research. 218. 418-423 (1995)
[文献書誌] Yamamoto, H. et al.: "A←brogation of lung metastasis of human fibrosarcoma cells by ribozyme-mediated suppression of integrin α6 subunit expression" International Journal of Cancer. (in press). (1996)
[文献書誌] Obara, M. et al.: "Possible involvement of the α5 subunit of α5β1 integrin with the synergistic region of the central cell-binding domain of fibronectin in cells to fibronectin binding." Exp. Cell Res.216. 273-276 (1995)
[文献書誌] 坂倉照よ・小原政信: "細胞外マトリックスの接着および反接着作用" 医学のあゆみ. 174. 32-35 (1995)
[文献書誌] Itano, N. et al.: "Phosphorylation of membrane-intercalated proteoglycan, syndecan-2, expressed in a stroma-inducing clone frome a mouse Lewis lung carcinoma" Biochemical Journal. (in press). (1996)
[文献書誌] Yano, H. et al.: "Transcriptional regulation of the chicken caldesmon gene: activation of gizzard type caldesmon promoter a CArG box-like motif" Journal of Biological Chemistry. 270. 23661-23666 (1995)
[文献書誌] Yoshida, K. et al.: "Reperfusion of rat heart after brief ischemia induces proteolysis of calspectin (Non-erythroid spectrin or fodrin) by calpain." Circulation Res.77. 603-610 (1995)
[文献書誌] Ilic D. et al.: "Reduced cell motility and enhanced focal adhesion contact formation in cells from FAK-dedicient mice." Nature. 377. 539-544 (1995)
[文献書誌] Ozawa, K. et al.: "Translocation of cortactin (p80/85) to the actin-based cytoskeleton during thrombin receptor-mediated platelet activation" Exp. Cell Res.221. 197-204 (1995)
[文献書誌] Ozawa, K. et al.: "Phase specific association of heterotrimeric GTP binding proteins with the actin-based cytoskeleton during thrombin receptor-mediated platelet activation" FEBS Lett.(in press). (1996)
[文献書誌] Saga, H. et al.: "Paradoxical expression of α-smooth muscle cells with their phenotype-dependent manner" EMBO J.(in press). (1996)
[文献書誌] Ogawara, et al.: "Differential targeting of protein kinase C and CaM kinase II signalings to vimentin" Journal of Cell Biology. 109. 1066 (1995)
[文献書誌] Sekimata, et al.: "Detection of protein kinase activity specifically activated at metaphase-anaphase transition" Journal of Cell Biology. (in press). (1996)
[文献書誌] Takai, et al.: "Mitosis-specific phosphorylation of vimentin by protein kinase C coupled with reorganization of intracellular membranes." Journal of Cell Biology. (in press). (1996)
[文献書誌] Kumagai, N. et al.: "Lysophosphatidic acid-induced activation of protein Ser/Thr kinases in cultured rat 3Y1 fibroblasts possible involvement in rho p21- mediated signalling." FEBS. Lett.366. 11 (1995)
[文献書誌] Saito Y. et al.: "Identification of Glu^<173> as the critical amino acid residue for the ADP-ribosyltransferase activity of Clostridium botulinum C3 exoenzme." FEBS. Lett.371. 105 (1995)
[文献書誌] Seckl M. J., et al.: "Guanosine 5'-3-O-(Thio) triphosphate stimulates tyrosine phosphorylation of p125^<FAK> and paxillin in permeabilized Swiss 3T3 cells" Journal of Biological Chemistry. 270. 6984-6990 (1995)
[文献書誌] Madalue, P. et al.: "A←novel partner for the GTP-bound forms of rho and rac" FEBS. Lett.377. 243-248 (1995)
[文献書誌] Watanabe, G. et al.: "Protein Kinase, PKN, and PKN-Related protein, rhophilin, as targets of small GTPasse rho" Science. (in press). (1995)
[文献書誌] Ishizaki, T. et al.: "The small GTP-binding protein rho binds to and activates a 160 kDa Ser/Thr protein kinase homologous to myotonic dystrophy kinase" EMBO.Journal. (in press). (1996)
[文献書誌] Maeda, N. et al.: "Purification, characterization and developmental expression of a brain-specific chondroitin sulfate proteoglycan, 6B4 proteoglycan/phosphacan" Neuroscience. 67. 23-35 (1995)
[文献書誌] Maeda, N. et al.: "6B4 proteoglycan/phosphacan is a repulsive substrate but promotes morphological differentiation of cortical neurons" Development. 122. 647-658 (1995)
[文献書誌] Takeda, H. et al.: "V-src kinase shifts the cadherin based cell adhesion from the strong to the weak state and β-catenin is not required for the shift" Journal of Cell Biology. 131. 1839-1847 (1995)
[文献書誌] Ando Akatuka, Y. et al.: "Interspecies diversity of the occludin sequence: cDNA cloning of human, mouse, dog and rat-kangaroo homologues" Journal of Cell Biology. (in press). (1996)

1995 年度 実績報告書

抗-細胞接着の分子機構と生理機能の解析

研究代表者

木全 弘治 愛知医科大学, 分子医科学研究所, 教授 (10022641)

研究成果

[文献書誌] Ito, K. et al.: "Multiple forms of mouse PG-M, a large chondroitin sufate proteoglycan generated by alternative splicing" The Journal of Biological Chemistry. 270. 958-965 (1995)

[文献書誌] Zako, M. et al.: "Expression of PG-M(V3), an alternatively spliced form of PG-M without a chondroitin sulfate attachment region in mouse and human tissue." The Journal of Biological Chemistry. 270. 3914-3918 (1995)

[文献書誌] Shinomura, T. et al.: "The gene structure and organization for mouse PG-M, a large chondroitin sulfate proteoglycan: Genomic background for the generation of multiple PG-M transcripts" The Journal of Biological Chemistry. 270. 10328-10333 (1995)

[文献書誌] Kato, S. et. al.: "Chondroitin sulfate immobilized onto culture substrates modulates 6DNA synthesis, tyrosine aminotransferase induction, and intercellular communication in primary rat hepatocytes" Cell Structure and function. 20. 199-209 (1995)

[文献書誌] Fukuoka, M. et al.: "Molecular cloning and expression of chick chondrocyte chondroitin 6-sulfotransferase" The Journal of Biological Chemistry. 270. 18575-18580 (1995)

[文献書誌] Nagai, N. et al.: "Quercetin suppresses heat shock response by down regulation of HSP1" Biochemical and Biophysical Research Communications. 208. 1099-1105 (1995)

[文献書誌] Higashi, T. et al.: "Activation of heat shock factor 1 in rat brain during cerebral ischemia or after heat shock" Mol. Brain Res.34. 262-270 (1995)

[文献書誌] Nakai A. et al.: "The DNA-Binding properties of two heat shock factors, HSF1 and HSF3, are induced in the avian erythroblast cell line HD6" Molecular and Cellular Biology. 15. 5268-5278 (1995)

[文献書誌] Yasuda, K. et al.: "Cloning and expression of murine high-molecular-weight heat shock proteins, HSP105" The Journal of Biological Chemistry. 270. 29718-29723 (1995)

[文献書誌] Nagata, K. et al.: "HSP47: a collagen-specific molecular chaperone in the endoplasmic reticulum" Trends in Biochemical Sciences. 21. 23-26 (1996)

[文献書誌] Satoh, M. et al.: "Intracellular interaction of collagen-specific stress protein HSP47 with newly synthesized procollagen" The Journal of Cell Biology. (in press). (1996)

[文献書誌] Jeon, H. et al.: "Pericytes from microvessel fragment produce type IV collagen and multiple laminin isoforms" Biosci. Biotech. Biochem.(in press). (1995)

[文献書誌] Kumagai, C. et al.: "Laminin α3-antiserum recognizes α chain in laminin 5 (kalinnin) but not that in laminin 6 (K-laminin) of human keratinocytes" Biochim. Biophys. Acta. (in press). (1995)

[文献書誌] Watanabe, E. et al.: "Distribution of brain-specific proteoglycan, neurocan, and the correspondion mRNA during the formation of barrels in the rat" European Journal of Neuroscience. 7. 547-554 (1995)

[文献書誌] Katoh-Semba, R. et al.: "Chondroitin sulfate proteoglycan in the rat brain: Candidates for axon barriers of sensory neurons and the possible" Nerosci. Let.189. 25-28 (1995)

[文献書誌] Maeda, N. et al.: "Purfication, characterization and developmental expression of brain-specific chondroitin sulfate-" Neurosci.67. 23-35 (1995)

[文献書誌] Watanabe, E. et al.: "Neuroglycan C, a novel membrane-spanning chondroitin sulfate proteoglycan that is restricted to the brain" Journal of Biological Chemistry. 270. 26876-26882 (1995)

[文献書誌] 大平 敦彦: "神経回路網形成期の脳に発現しているプロテオグリカンの構造と機能" 日本神経精神薬理学雑誌. 171-176 (1995)

[文献書誌] Mizutani, H. et al.: "Nodular scleroderma: Focally increased tenascin expession differing from that in the surrounding scleroderma soin" J. Dermatology. 22. 267-271 (1995)

[文献書誌] Ishihara, A. et al.: "Tenascin expressuin in cancer cells and stroma of human breast cancer amd its prognostic significance" Clin. Cancer Res.1. 1035-1041 (1995)

[文献書誌] Yoshida, T. et al.: "Tenascin expression in breast cancer cell: A maker for poor grognosis in breast cancer" Cancer Letter. 90. 65-73 (1995)

[文献書誌] Ichihara-Tanaka, K. et al.: "Role of the carboxyl-terminal Fib2 domain in fibronectin matrix assembly" Journal of Cell Science. 108. 907-915 (1995)

[文献書誌] Healy J. M. et al.: "Peptide ligands for integrin αvβ3 selected from random phage display libraries" Biochemistry. 34. 3948-3955 (1995)

[文献書誌] Ohashi, H. et al.: "Up-regulation of integrin α5β1 expression by interleukin-6 in rabbit corneal epithelial cells" Experimental Cell Research. 218. 418-423 (1995)

[文献書誌] Yamamoto, H. et al.: "A←brogation of lung metastasis of human fibrosarcoma cells by ribozyme-mediated suppression of integrin α6 subunit expression" International Journal of Cancer. (in press). (1996)

[文献書誌] Obara, M. et al.: "Possible involvement of the α5 subunit of α5β1 integrin with the synergistic region of the central cell-binding domain of fibronectin in cells to fibronectin binding." Exp. Cell Res.216. 273-276 (1995)

[文献書誌] 坂倉照よ・小原政信: "細胞外マトリックスの接着および反接着作用" 医学のあゆみ. 174. 32-35 (1995)

[文献書誌] Itano, N. et al.: "Phosphorylation of membrane-intercalated proteoglycan, syndecan-2, expressed in a stroma-inducing clone frome a mouse Lewis lung carcinoma" Biochemical Journal. (in press). (1996)

[文献書誌] Yano, H. et al.: "Transcriptional regulation of the chicken caldesmon gene: activation of gizzard type caldesmon promoter a CArG box-like motif" Journal of Biological Chemistry. 270. 23661-23666 (1995)

[文献書誌] Yoshida, K. et al.: "Reperfusion of rat heart after brief ischemia induces proteolysis of calspectin (Non-erythroid spectrin or fodrin) by calpain." Circulation Res.77. 603-610 (1995)

[文献書誌] Ilic D. et al.: "Reduced cell motility and enhanced focal adhesion contact formation in cells from FAK-dedicient mice." Nature. 377. 539-544 (1995)

[文献書誌] Ozawa, K. et al.: "Translocation of cortactin (p80/85) to the actin-based cytoskeleton during thrombin receptor-mediated platelet activation" Exp. Cell Res.221. 197-204 (1995)

[文献書誌] Ozawa, K. et al.: "Phase specific association of heterotrimeric GTP binding proteins with the actin-based cytoskeleton during thrombin receptor-mediated platelet activation" FEBS Lett.(in press). (1996)

[文献書誌] Saga, H. et al.: "Paradoxical expression of α-smooth muscle cells with their phenotype-dependent manner" EMBO J.(in press). (1996)

[文献書誌] Ogawara, et al.: "Differential targeting of protein kinase C and CaM kinase II signalings to vimentin" Journal of Cell Biology. 109. 1066 (1995)

[文献書誌] Sekimata, et al.: "Detection of protein kinase activity specifically activated at metaphase-anaphase transition" Journal of Cell Biology. (in press). (1996)

[文献書誌] Takai, et al.: "Mitosis-specific phosphorylation of vimentin by protein kinase C coupled with reorganization of intracellular membranes." Journal of Cell Biology. (in press). (1996)

[文献書誌] Kumagai, N. et al.: "Lysophosphatidic acid-induced activation of protein Ser/Thr kinases in cultured rat 3Y1 fibroblasts possible involvement in rho p21- mediated signalling." FEBS. Lett.366. 11 (1995)

[文献書誌] Saito Y. et al.: "Identification of Glu^<173> as the critical amino acid residue for the ADP-ribosyltransferase activity of Clostridium botulinum C3 exoenzme." FEBS. Lett.371. 105 (1995)

[文献書誌] Seckl M. J., et al.: "Guanosine 5'-3-O-(Thio) triphosphate stimulates tyrosine phosphorylation of p125^<FAK> and paxillin in permeabilized Swiss 3T3 cells" Journal of Biological Chemistry. 270. 6984-6990 (1995)

[文献書誌] Madalue, P. et al.: "A←novel partner for the GTP-bound forms of rho and rac" FEBS. Lett.377. 243-248 (1995)

[文献書誌] Watanabe, G. et al.: "Protein Kinase, PKN, and PKN-Related protein, rhophilin, as targets of small GTPasse rho" Science. (in press). (1995)

[文献書誌] Ishizaki, T. et al.: "The small GTP-binding protein rho binds to and activates a 160 kDa Ser/Thr protein kinase homologous to myotonic dystrophy kinase" EMBO.Journal. (in press). (1996)

[文献書誌] Maeda, N. et al.: "Purification, characterization and developmental expression of a brain-specific chondroitin sulfate proteoglycan, 6B4 proteoglycan/phosphacan" Neuroscience. 67. 23-35 (1995)

[文献書誌] Maeda, N. et al.: "6B4 proteoglycan/phosphacan is a repulsive substrate but promotes morphological differentiation of cortical neurons" Development. 122. 647-658 (1995)

[文献書誌] Takeda, H. et al.: "V-src kinase shifts the cadherin based cell adhesion from the strong to the weak state and β-catenin is not required for the shift" Journal of Cell Biology. 131. 1839-1847 (1995)

[文献書誌] Ando Akatuka, Y. et al.: "Interspecies diversity of the occludin sequence: cDNA cloning of human, mouse, dog and rat-kangaroo homologues" Journal of Cell Biology. (in press). (1996)

1995 年度実績報告書

木全弘治愛知医科大学, 分子医科学研究所, 教授 (10022641)

[文献書誌] 大平敦彦: "神経回路網形成期の脳に発現しているプロテオグリカンの構造と機能" 日本神経精神薬理学雑誌. 171-176 (1995)