Desensitization, internalization, and signaling functions of β-arrestins demonstrated by RNA interference
Desensitization, internalization, and signaling functions of β-arrestins demonstrated by RNA interference
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DOI：
10.1073/pnas.262789099
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发表时间：
2003-02-18
影响因子：
11.1
通讯作者：
Lefkowitz, RJ
中科院分区：
文献类型：
--
作者：
Ahn, S;Nelson, CD;Lefkowitz, RJ
β-抑制蛋白与活化的G蛋白偶联受体激酶磷酸化受体结合，这导致它们对G蛋白的脱敏、通过网格蛋白包被的小凹的内化和通过越来越多的“支架”途径的信号传导。为了促进新的适配器和β-arrestins的信号作用的发现，我们已经开发并验证了一种普遍适用的干扰RNA方法，用于选择性地抑制β-arrestins 1或2的表达高达95%。HEK 293细胞中β-抑制蛋白的耗竭导致响应于β（2）-肾上腺素能受体刺激的cAMP生成增强，β（2）-肾上腺素能受体和血管紧张素II受体内化显著降低，以及血管紧张素II对MAP激酶ERK 1和2的活化受损。这种方法应该允许在许多七跨膜受体系统中发现β-arrestins的新信号和调节作用。
beta-Arrestins bind to activated G protein-coupled receptor kinase-phosphorylated receptors, which leads to their desensitization with respect to G proteins, internalization via clathrin-coated pits, and signaling via a growing list of "scaffolded" pathways. To facilitate the discovery of novel adaptor and signaling roles of beta-arrestins, we have developed and validated a generally applicable interfering RNA approach for selectively suppressing beta-arrestins 1 or 2 expression by up to 95%. beta-Arrestin depletion in HEK293 cells leads to enhanced cAMP generation in response to beta(2)-adrenergic receptor stimulation, markedly reduced beta(2)-adrenergic receptor and angiotensin II receptor internalization and impaired activation of the MAP kinases ERK 1 and 2 by angiotensin II. This approach should allow discovery of novel signaling and regulatory roles for the beta-arrestins in many seven-membrane-spanning receptor systems.