Nonmuscle Myosin II and Upstream and Downstream Signaling

非肌肉肌球蛋白 II 和上下游信号传导

• 批准号: 8557932
• 负责人: Robert Adelstein
• 金额: $ 42.52万
• 依托单位: NATIONAL HEART, LUNG, AND BLOOD INSTITUTE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8557932
• 关键词: Attenuated; Cell Communication; Cell Line; Cells; Co-Immunoprecipitations; Epidermal Growth Factor Receptor; Ligands; Light; Mass Spectrum Analysis; Motor; Myosin Type II; Phosphotransferases; Play; Process; Proteins; Proto-Oncogene Proteins c-akt; Role; Signal Pathway; Signal Transduction; Small Interfering RNA; blebbistatin; cell type; mutant; non-muscle myosin; paralogous gene; receptor-mediated signaling

Ligand-induced internalization of the epidermal growth factor receptor (EGFR) is an important process for regulating signal transduction, cellular dynamics, and cell-cell communication. We have recently demonstrated that nonmuscle myosin II (NM II) is required for the internalization of the EGFR and to trigger the EGFR-dependent activation of ERK and AKT. The EGFR was identified as a protein that interacts with NM II by co-immunoprecipitation and mass spectrometry analysis. This interaction requires both the regulatory light chain 20 (RLC20) of NM II and the kinase domain of the EGFR. Two paralogs of NM II, NM II-A and NM II-B can act to internalize the EGFR, depending on the cell type and paralog content of the cell line. Loss (siRNA) or inhibition (25M blebbistatin) of NM II attenuates the internalization of the EGFR and impairs EGFR-dependent activation of ERK and AKT. Both internalization of the EGFR and downstream signaling to ERK and AKT can be partially restored in siRNA-treated cells by introduction of wild type (WT) GFP-NM II, but cannot be restored by motor mutant NM II. Taken together, these results suggest that NM II plays a role in the internalization of the EGFR and EGFR-mediated signaling pathways.

配体诱导的表皮生长因子受体 (EGFR) 内化是调节信号转导、细胞动力学和细胞间通讯的重要过程。我们最近证明非肌肉肌球蛋白 II (NM II) 是 EGFR 内化和触发 EGFR 依赖性 ERK 和 AKT 激活所必需的。通过免疫共沉淀和质谱分析，EGFR 被鉴定为与 NM II 相互作用的蛋白质。这种相互作用需要 NM II 的调节轻链 20 (RLC20) 和 EGFR 的激酶结构域。 NM II 的两种旁系同源物、NM II-A 和 NM II-B 可以内化 EGFR，具体取决于细胞系的细胞类型和旁系同源物含量。 NM II 的缺失 (siRNA) 或抑制 (25M blebbistatin) 会减弱 EGFR 的内化并损害 EGFR 依赖性的 ERK 和 AKT 激活。通过引入野生型 (WT) GFP-NM II，EGFR 的内化以及 ERK 和 AKT 的下游信号传导可以在 siRNA 处理的细胞中部分恢复，但不能通过运动突变体 NM II 恢复。综上所述，这些结果表明 NM II 在 EGFR 内化和 EGFR 介导的信号通路中发挥作用。