Genome-wide screen of gamma-secretase-mediated intramembrane cleavage of receptor tyrosine kinases
Genome-wide screen of gamma-secretase-mediated intramembrane cleavage of receptor tyrosine kinases
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DOI：
10.1091/mbc.e17-04-0261
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发表时间：
2017-11-01
影响因子：
3.3
通讯作者：
Elenius, Klaus
中科院分区：
文献类型：
--
作者：
Merilahti, Johannes A. M.;Ojala, Veera K.;Elenius, Klaus
受体酪氨酸激酶 (RTK) 已被证明通过调节的膜内蛋白水解发出信号，其中胞外域脱落和随后由 γ 分泌酶进行的膜内裂解导致释放具有功能活性的可溶性细胞内受体片段。然而，对于大多数 RTK 来说，尚不清楚它们是否可以利用这种新的信号机制。在这里，我们使用系统范围的屏幕来解决人类 RTK 中对 γ 分泌酶裂解的敏感性频率。筛选覆盖了 55 个人类 RTK 中的 45 个，识别出 12 个新的以及所有 9 个先前发表的 γ 分泌酶底物。我们通过证明内源性 AXL 中可溶性细胞内片段的释放依赖于脱落酶解整合素和金属蛋白酶 10 (ADAM10) 以及 γ 分泌酶成分 presenilin-1，对筛选进行了生化验证。可裂解 RTK 的功能分析表明，TAM 家族成员 AXL 或 TYRO3 过表达所促进的增殖取决于 γ 分泌酶裂解。综上所述，这些数据表明，γ-分泌酶介导的裂解为许多人类 RTK 提供了额外的信号传导机制。
Receptor tyrosine kinases (RTKs) have been demonstrated to signal via regulated intramembrane proteolysis, in which ectodomain shedding and subsequent intramembrane cleavage by gamma-secretase leads to release of a soluble intracellular receptor fragment with functional activity. For most RTKs, however, it is unknown whether they can exploit this new signaling mechanism. Here we used a system-wide screen to address the frequency of susceptibility to gamma-secretase cleavage among human RTKs. The screen covering 45 of the 55 human RTKs identified 12 new as well as all nine previously published gamma-secretase substrates. We biochemically validated the screen by demonstrating that the release of a soluble intracellular fragment from endogenous AXL was dependent on the sheddase disintegrin and metalloprotease 10 (ADAM10) and the gamma-secretase component presenilin-1. Functional analysis of the cleavable RTKs indicated that proliferation promoted by overexpression of the TAM family members AXL or TYRO3 depends on gamma-secretase cleavage. Taken together, these data indicate that gamma-secretase-mediated cleavage provides an additional signaling mechanism for numerous human RTKs.