Roles of phosphorylation of ERM proteins and ERM-binding membrane proteins in cellular morphogenesis

ERM 蛋白和 ERM 结合膜蛋白磷酸化在细胞形态发生中的作用

• 批准号: 11670117
• 负责人: YONEMURA Shigenobu
• 金额: $ 2.43万
• 依托单位: KYOTO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11670117/
• 关键词: ERM proteins; ERM-binding membrane proteins; microvilli; phosphorylation; TCA fixation; Rho; PIP2; neomycin; ERM; C3; 形態形成; 脱リン酸化; トリクロロ酢酸

1 : Effects of overexpression of ERM-binding membrane proteins (CD43, CD44 and ICAM-2) on microvilli formationOverexpression of these membrane proteins in L and CV1 cells resulted in elongation of microvilli, indicating these proteins are major components of microvilli together with ERM proteins.2 : Localization of ERM proteins phosphorylated at the COOH-terminal threonine residue (CPERMs)We developed new fixation protocol using trichloroacetic acid as a fixative to determine precise localization of CPERMs. They were localized just beneath the plasma membrane of microvilli, reflecting that CPERMs are activated ERM proteins as membrane-cytoskeleton cross-linkers.3 : ERM protein activation in cellular morphogenesisUse of Rho inhibitor, C3, protein kinase inhibitor, staurosporine, and PIP2 inhibitor, neomycin revealed that importance of Rho activity and phosphorylation of ERM proteins depends on cell type. However PIP2 was found to be essential without exception for activation of ERM proteins as well as microvilli formation.

一曰：ERM-binding membrane proteins过度表达的影响（CD 43、CD 44和ICAM-2）对微绒毛形成的影响这些膜蛋白在L和CV 1细胞中的过表达导致微绒毛的延长，表明这些蛋白与ERM蛋白一起是微绒毛的主要成分。2：在COOH末端苏氨酸残基磷酸化的ERM蛋白（CPERMs）的定位我们开发了新的固定方案，使用三氯乙酸作为固定剂，以确定精确定位的CPERMs。3：ERM蛋白在细胞形态发生中的活化使用Rho抑制剂C3、蛋白激酶抑制剂staurosporine和PIP 2抑制剂新霉素揭示了Rho活性和ERM蛋白磷酸化的重要性取决于细胞类型。然而，发现PIP 2对于ERM蛋白的活化以及微绒毛的形成无一例外地是必需的。

项目成果

月田承一郎 他、: "岩波講座・現代医学の基礎 分子・細胞の生物学II 第6章 細胞骨格と細胞運動"岩波書店. 17 (2000)

Joichiro Tsukita等人：“岩波讲座：现代医学分子和细胞生物学基础II第6章细胞骨架和细胞运动”岩波书店17（2000）。

Yoshinori Doi: "Normal development of mice and unimpaired cell adhesion/cell Motility/Actin-based cytoskeleton without compasatory・・・"J.Biological Chemistry. 274. 2315-2321 (1999)

Yoshinori Doi：“小鼠的正常发育和未受损的细胞粘附/细胞运动/基于肌动蛋白的细胞骨架，无需补偿......”J.Biological Chemistry 274. 2315-2321 (1999)

Noriyuki Sonoda: "Clostridium perfrongens enterotoxin fragment removes specific claudins from tight junction strands : Evidence for direct involvement・・・"J.Cell Biology. 147. 195-204 (1999)

Noriyuki Sonoda：“产气荚膜梭菌肠毒素片段从紧密连接链中去除特定的密蛋白：直接参与的证据......”J.Cell Biology 147. 195-204 (1999)

S.Yonemura, Sa.Tsukita and Sh.Tsukita: "Direct involvement of ezrin/radixin/moesin (ERM) -binding membrane proteins in the organization of microvilli in collaboration with activated ERM proteins."J.Cell Biol.. 145. 1497-1509 (1999)

S.Yonemura、Sa.Tsukita 和 Sh.Tsukita：“ezrin/radixin/moesin (ERM) 结合膜蛋白与激活的 ERM 蛋白协同参与微绒毛组织的直接参与。”J.Cell Biol.. 145. 1497

N.Sonoda, M.Furuse, H.Sasaki, S.Yonemura, J.Katahira, Y.Horiguchi and Sh.Tsukita: "Clostridium perfringens enterotoxin fragment removes specific claudins from tight strands : evidence for direct involvement of claudins in tight junction junction barrier."

N.Sonoda、M.Furuse、H.Sasaki、S.Yonemura、J.Katahira、Y.Horiguchi 和 Sh.Tsukita：“产气荚膜梭菌肠毒素片段从紧密链中去除特定的密蛋白：密蛋白直接参与紧密连接的证据