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Adaptor protein APPL1 interacts with EGFR to orchestrate EGF-stimulated signaling

  • Article
  • Life & Medical Sciences
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Science Bulletin

Abstract

Epidermal growth factor receptor (EGFR) mediates multiple signaling pathways that regulate cell migration, proliferation, and differentiation. Adaptor protein APPL1 has been reported to function as a downstream effector of EGFR signaling pathway. However, molecular mechanisms underlying the role of APPL1 downstream of EGFR signaling remains elusive. Here, we identified APPL1 as a critical molecule that interacts with EGFR. Suppression of APPL1 by siRNA inhibited EGF-stimulated Akt phosphorylation. Functionally, EGF stimulation of cells caused phosphorylation of APPL1 at Ser636, which subsequently promoted the interaction between APPL1 and EGFR, indicating that APPL1 sensitizes EGF stimulation by acting at a site downstream of the EGFR signaling. Importantly, non-phosphorylatable mutant of APPL1 reduced cell migration compared with wild-type APPL1 in an Akt-dependent manner. Our study reveals a novel function of APPL1 in EGF signaling and defines a novel molecular mechanism by which phosphorylation of APPL1 upon EGF stimulation regulates cell migration underlying EGF-stimulated Akt pathway.

摘要

表皮生长因子信号轴调控细胞可塑性与命运决定等重要 细胞生理学过程, 但细胞信号转导枢纽蛋白APPL1 如何介导表皮生 长因子信号轴在细胞迁移过程中的分子机制仍不甚清晰。我们在本 项研究中鉴定出APPL1 是一个新的表皮生长因子受体结合蛋白并调 控表皮生长因子介导的细胞定向迁移活动。利用RNA 干扰技术, 我 们发现:在细胞中敲低APPL1 可下调EGF 介导的Akt 和GSK3β 的 磷酸化水平。进一步的解析发现, EGF 刺激会引起APPL1 第636 位丝氨酸的磷酸化, 进而增强APPL1 与表皮生长因子受体的结合 力。通过转入模拟磷酸化及不可被磷酸化的APPL1 突变体, 我们发 现抑制APPL1 第636 位丝氨酸的磷酸化可阻碍表皮生长因子介导 的细胞迁移活动。我们认为, APPL1 蛋白通过第636 位丝氨酸的磷 酸化增强其与表皮生长因子受体的结合及其在细胞质膜上的稳定性, 从而延续表皮生长因子信号轴的活性并有效调控细胞的定向迁移活动。

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Acknowledgments

We are grateful to Xuebiao Yao (University of Science and Technology of China) and Donald L. Hill (University of Alabama at Birmingham) for support. This work is, in whole or in part, supported by the National Natural Science Foundation of China (31501130, 31501095), China Postdoctoral Science Foundation (2014M560517) and Anhui Provincial Natural Science Foundation (1508085SMC213).

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    Authors and Affiliations

    1. Anhui Key Laboratory of Cellular Dynamics and Chemical Biology, University of Science and Technology of China, Hefei, 230027, China

      Jiajia Zhou, Hongsheng Liu, Shu Zhou & Xing Liu

    2. Guangzhou Women and Children’s Medical Center, Guangzhou, 510623, China

      Ping He

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    Correspondence to Xing Liu.

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    Author contributions

    J.Z., H.L. and X.L. conceived the project; J.Z., S.Z. and P.H. performed biochemical and cell biological experiments; J.Z., S.Z. and X.L. performed data analyses; J.Z., H.L., P.H., and X.L. wrote and edited the manuscript.

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    The authors declare that they have no conflict of interest.

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    SPECIAL TOPIC: Lipid Metabolism and Human Metabolic Disorder.

    J. Zhou and H. Liu contributed equally to this work.

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    Zhou, J., Liu, H., Zhou, S. et al. Adaptor protein APPL1 interacts with EGFR to orchestrate EGF-stimulated signaling. Sci. Bull. 61, 1504–1512 (2016). https://doi.org/10.1007/s11434-016-1157-0

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    • DOI: https://doi.org/10.1007/s11434-016-1157-0

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