The roles of cofilin and LIM-kinase in chemotaxis of phagocytes

• Project/Area Number: 15590090
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Biological pharmacy
• Research Institution: National Institute of Health Sciences
• Principal Investigator: SUZUKI Kazuhiro NIHS, Division of Biosignaling, Section chief, 代謝生化学部, 室長 (10154527)
• Co-Investigator(Kenkyū-buntansha): ADACHI Reiko NIHS, Division of Biosignaling, Senior researcher, 代謝生化学部, 主任研究官 (10291113)
• Project Period (FY): 2003 – 2004
• Project Status: Completed (Fiscal Year 2004)
• Budget Amount: ¥3,100,000 (Direct Cost: ¥3,100,000); Fiscal Year 2004: ¥1,500,000 (Direct Cost: ¥1,500,000); Fiscal Year 2003: ¥1,600,000 (Direct Cost: ¥1,600,000)
• Keywords: neutrophil / HL-60 cell / chemotaxis / cofilin / phosphorylation / Interleukin-8 / PI3-kinase / siRNA / 食細胞 / siRNA

Research Abstract

Cofilin is a ubiquitous actin-binding protein. Only unphosphorylated cofilin binds actin and severs or depolymerizes a filamentous actin(F-actin) and cofilin is phosphrylated by LIM-kinase, a specific kinase of cofilin, to be an inactive form which does not bind actin. The phosphorylated cofilin is dephosphorylated by slingshot, a specific phosphatase, to be an active form. Recently we have reported that cofilin plays regulatory roles in superoxide production and phagocytosis by phagocytes. In this study, the roles of cofilin hi chemotaxis of leukocytes were investigated. Interleukin 8(IL-8), a potent physiological chemokine, triggered quick dephoshorylation and subsequent rephosphorylation of cofilin. The phosphorylation of cofilin was inhibited by S3-R peptide which consisted of a membrane-permeable peptide and a peptide of phosphorylation site of cofilin. When the S3-R peptide was introduced into neutrophil-like HL-60 cells, the chemotactic activity was enhanced while control peptide which contained an inverted sequence of phosphorylation site of cofilin did not such an enhancing effect. Wortmannin and LY294002,inhibitors of PI3 kinase, suppressed the chemotaxis and phosphorylation turnover of cofilin. On the other hand, siRNA of cofilin caused down-regulation of cofilin and inhibited the chemotaxis. Based on these results it is suggested that unphosphrylated active cofilin plays a critical role in the chemtaxis of phagocytes and PI3-kinase is involved in the control of phosphorylation/dephosphorylation cycle of cofilin.

Research Products

• [Journal Article] Two HLA-E Mutants Segregate the Inhibitory NK Receptor CD94/NKG2A and the Activating Receptor CD94/NKG2C Both of Which Bind the Top of HLA-E (2004)
  • Author(s): Wada, H., Matsumoto, N., Maenaka, K., Suzuki, K., Yamamoto, K
  • Journal Title: Euro.J.Immunol. 34 Pages: 81-90
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Augmentation of NK cell-mediated cytotoxicity to tumor cells by inhibitory NK cell receptor blockers (2004)
  • Author(s): Tajima, K., Matsumoto, N., Ohmori, K., Wada, H., Ito, M., Suzuki, K., Yamamoto, K.
  • Journal Title: Int.Immunol. 16 Pages: 385-393
  • NAID: 10013084017
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Ribosomal protein S18 identified as a cofilin-binding protein by using phage display library (2004)
  • Author(s): Kusui, K., Sasaki, H., Adachi, R., Matsui, S., Yamamoto, K., Yamaguchi, T., Kasahara, T., Suzuki, K.
  • Journal Title: Mol.Cell.Biochem. 262 Pages: 187-193
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Ribosomal protein S18 identified as a cofilin-binding protein by using phage display library (2004)
  • Author(s): Kusui, K., Sasaki, H., Adachi, R., Matsui, S., Yamamoto, K., Yamaguchi, T., Kasahara, T., Suzuki.K.
  • Journal Title: Mol.Cell.Biochem. 262 Pages: 187-193
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] The inhibitory NK cell receptor CD94/NKG2A and the activating receptor CD94/NKG2C bind the Top of HLA-E through mostly shared but partly distinct sets of HLA-E residues (2004)
  • Author(s): Wada, H., Matsumoto, N., Maenaka, K., Suzuki.K., Yamamoto, K
  • Journal Title: Eur.J.Immunol. 34 Pages: 81-90
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Augmentation of NK cell-mediated cytotoxicity to tumor cells by inhibitory NK cell receptor blockers. (2004)
  • Author(s): Tajima, K., Matsumoto, N., Ohmori, K., Wada, H., Ito, M., Suzuki.K., Yamamoto, K.
  • Journal Title: Int.Immunol. 16 Pages: 385-393
  • NAID: 10013084017
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Two HLA-E Mutants Segregate the Inhibitory NK Receptor CD94/NKG2A and the Activating Receptor CD94/NKG (2004)
  • Author(s): Wada, H., Matsumoto, N., Maenaka, K., Suzuki, K., Yamamoto, K
  • Journal Title: Euro.J.Immunol. 34 Pages: 81-90
• [Journal Article] Triphenyltin enhances the neutrophilic differentiation of promyelocytic HL-60 (2003)
  • Author(s): Watanabe, H., Adachi, R., Hirayama, A., Kasahara, T., Suzuki, K
  • Journal Title: Biochem.Biophys.Res.Commun. 306 Pages: 26-31
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Bisphenol A significantly enhances the neutrohilic differentiation of promyelocytic HL-60 cells (2003)
  • Author(s): Watanabe, H., Adachi, R., Kusui, K., Hirayama, A., Kasahara, T., Suzuki, K
  • Journal Title: Int.Immunopharmacol. 3 Pages: 1601-1608
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] 食細胞の機能発現とLIMキナーゼ-コフィリンによるアクチン細胞骨格制御 (2003)
  • Author(s): 安達玲子, 鈴木和博
  • Journal Title: 生化学 75 Pages: 1238-1243
  • NAID: 50000316593
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Possible Involvement of Optimally Phosphorylated L-Plastin in Activation of Superoxide-Generating (2003)
  • Author(s): Oshizawa, T., Yamaguchi, T., Suzuki, K., Yamamoto, Y., Hayakawa, T
  • Journal Title: J.Biochem. 134 Pages: 827-834
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Possible Involvement of Optimally Phosphorylated L-Plastin in Activation of Superoxide-Generating NADPH Oxidase. (2003)
  • Author(s): Oshizawa, T., Yamaguchi, T., Suzuki, K., Yamamoto, Y., Hayakawa, T.
  • Journal Title: J.Biochem. 134 Pages: 827-834
  • NAID: 50000393691
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Relationship between phagocyte functions and regulation of actin cytoskeleton by LIM-kinase and cofilin (Review in Japanese) (2003)
  • Author(s): Adachi, R., Suzuki, K.
  • Journal Title: Seikagaku 75 Pages: 1238-1243
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Cryptococcus neoformans induces alterations in the cytoskeleton of human brain microvascular endotherial cells. (2003)
  • Author(s): Chen, S.H., Stins, M.F., Huang, S.H., Chen, Y.H., Kwon-Chung, K.J., Chang, Y., Kim, K.S., Suzuki, K., Jong, A.Y.
  • Journal Title: J.Med.Microbiol. 52 Pages: 961-970
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Bisphenol A significantly enhances the neutrohilic differentiation of prormyelocytic HL-60 cells (2003)
  • Author(s): Watanabe, H., Adachi, R., Kusui, K., Hirayama, A., Kasahara, T., Suzuki, K.
  • Journal Title: Int.Immunopharmacol. 3 Pages: 1601-1608
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Triphenyltin enhances the neutrophilic differentiation of promyelocytic HL-60 Cells (2003)
  • Author(s): Watanabe, H., Adachi, R., Hirayama, A., Kasahara, T., Suzuki, K.
  • Journal Title: Biochem.Biophys.Res.Commun. 306 Pages: 26-31
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Watanabe, H., Adachi, R., Hirayama, A., Kasahara, T., Suzuki, K: "Triphenyltin enhances the neutrophilic differentiation of promyelocytic HL-60"Biochem.Biophys.Res.Commun.. 306. 26-31 (2003)
• [Publications] Watanabe, H., Adachi, R., Kusui, K., Hirayama, A., Kasahara, T., Suzuki, K: "Bisphenol A significantly enhances the neutrohilic differentiation of promyelocytic HL-60 cells"Int.Immunopharmacol.. 3. 1601-1608 (2003)
• [Publications] 安達玲子, 鈴木和博: "食細胞の機能発現とLIMキナーゼ-コフィリンによるアクチン細胞骨格制御"生化学. 75. 1238-1243 (2003)
• [Publications] Oshizawa, T., Yamaguchi, T., Suzuki, K., Yamamoto, Y., Hayakawa, T: "Possible Involvement of Optimally Phosphorylated L-Plastin in Activation of Superoxide-Generating"J.Biochem.. 134. 827-834 (2003)
• [Publications] Wada, H., Matsumoto, N., Maenaka, K., Suzuki, K., Yamamoto, K: "Two HLA-E Mutants Segregate the Inhibitory NK Receptor CD94/NKG2A and the Activating Receptor CD94/NKG2C bind the top of HLA-E through mostly shared but partly distinct sets of HLA-E residues"Euro.J.Immunol.. 34. 81-90 (2004)
• [Publications] Tajima, K., Matsumoto, N., Ohmori, K., Wada, H., Ito, M., Suzuki, K., Yamamoto, K.: "Augmentation of NK cell-mediated cytotoxicity to tumor cells by inhibitory NK cell receptor blockers"Int.Immunol.. 16. 385-393 (2004)
• [Publications] Kusui, K., Sasaki, H., Adachi, R., Matsui, S., Yamamoto, K., Yamaguchi, T., Kasahara, T., Suzuki, K.: "Ribosomal protein S18 identified as a cofilin-binding protein by using phage display library"Mol.Cell.Biochem.. 2004(印刷中).