Cullin 3 regulates ADAMs-mediated ectodomain shedding of amphiregulin
Cullin 3 regulates ADAMs-mediated ectodomain shedding of amphiregulin
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Cullin 3 调节 ADAM 介导的双调蛋白胞外域脱落
DOI：
10.1016/j.bbrc.2018.03.097
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发表时间：
2018
影响因子：
3.1
通讯作者：
Higashiyama Shigeki
中科院分区：
文献类型：
--
作者：
Nakayama Hironao;Sakaue Tomohisa;Maekawa Masashi;Fujisaki Ayako;Higashiyama Shigeki
解整合素和金属蛋白酶（adintegrin and metalloproteinase，ADAM）家族是多种底物胞外区脱落的关键酶，参与多种生物学和病理学过程。然而，亚当斯的底物选择性的分子机制知之甚少。在这项研究中，我们观察到，破坏肌动蛋白聚合的药理学抑制剂，latrunculin A（LatA）和细胞松弛素D（CyD），诱导胞外域脱落的表皮生长因子（EGF）家族配体。由LatA或CyD诱导的脱落活性被金属蛋白酶抑制剂KB-R7785抑制，表明ADAMs介导的脱落受到肌动蛋白细胞骨架的严格控制。我们还研究了cullin家族，cullin-RING为基础的E3泛素连接酶的组成部分，在胞外域脱落的作用，因为cullin家族参与细胞骨架动力学的调节。通过特异性siRNA敲除cullin 3（Cul 3）可抑制EGF家族成员双调蛋白（AREG）的胞外域脱落，并且反应与RhoA GTPase的激活和应激纤维形成的诱导相关。另一方面，RhoA抑制剂C3转移酶挽救了Cul 3敲低减少的AREG脱落。这些结果描述了Cul 3通过以RhoA依赖性方式调节细胞骨架动力学来调节AREG脱落的新分子机制。
A disintegrin and metalloproteinase (ADAM) family are crucial enzymes for ectodomain shedding of multiple substrates and are involved in diverse biologic and pathologic processes. However, the molecular mechanism underlying substrate selectivity of ADAMs is poorly understood. In this study, we observed that disruption of actin polymerization by pharmacological inhibitors, latrunculin A (LatA) and cytochalasin D (CyD), induced ectodomain shedding of epidermal growth factor (EGF) family ligands. Induced shedding activity by LatA or CyD was suppressed by a metalloprotease inhibitor KB-R7785, indicating that ADAMs-mediated shedding is tightly controlled by actin cytoskeleton. We also investigated roles of cullin family, a component of cullin-RING based E3 ubiquitin ligases, in ectodomain shedding, since cullin family is implicated in the regulation of cytoskeletal dynamics. Knockdown of cullin 3 (Cul3) by a specific siRNA inhibited ectodomain shedding of amphiregulin (AREG), a member of EGF family, and responses were associated with activation of RhoA GTPase and induction of stress fiber formation. On the other hand, the RhoA inhibitor C3 transferase rescued AREG shedding reduced by Cul3 knockdown. These results describe a novel molecular mechanism of Cul3 to regulate AREG shedding by modulating cytoskeletal dynamics in a RhoA dependent manner.