Dopamine D2 and D3 receptors are linked to the actin cytoskeleton via interaction with filamin A
Dopamine D2 and D3 receptors are linked to the actin cytoskeleton via interaction with filamin A
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DOI：
10.1073/pnas.011538198
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发表时间：
2001-04-24
影响因子：
11.1
通讯作者：
Levenson, R
中科院分区：
文献类型：
--
作者：
Lin, RW;Karpa, K;Levenson, R
我们已经使用酵母双杂交的方法来揭示蛋白质的相互作用，涉及多巴胺受体的D2样亚家族。以多巴胺D_2S和D_3受体的第三环为诱饵，从人脑cDNA文库中筛选到一种与多巴胺D_2和D_3受体均相互作用的蛋白质--细丝蛋白A（FLN-A）。缺失作图定位多巴胺受体-FLN-A相互作用的N-末端段的D2和D3多巴胺受体和重复19 FLN-A。在培养的解离大鼠纹状体，FLN-A和D2受体共定位在整个神经元胞体和过程，以及在星形胶质细胞。在FLN-A缺陷的M2黑色素瘤细胞中D2多巴胺受体的表达导致D2受体主要位于细胞内，而在FLN-A重建的细胞中，D2受体主要位于质膜上。这些结果表明，FLN-A可能是D2多巴胺受体的适当细胞表面表达所必需的。D2和D3多巴胺受体与FLN-A的结合提供了一种机制，由此特异性多巴胺受体亚型可能通过肌动蛋白细胞骨架与下游信号传导组分功能性连接。
We have used a yeast two-hybrid approach to uncover protein interactions involving the D2-like subfamily of dopamine receptors. Using the third intracellular loop of the D2S and D3 dopamine receptors as bait to screen a human brain cDNA library, we identified filamin A (FLN-A) as a protein that interacts with both the D2 and D3 subtypes, The interaction with FLN-A was specific for the D2 and D3 receptors and was independently confirmed in pull-down and coimmunoprecipitation experiments. Deletion mapping localized the dopamine receptor-FLN-A interaction to the N-terminal segment of the D2 and D3 dopamine receptors and to repeat 19 of FLN-A, In cultures of dissociated rat striatum, FLN-A and D2 receptors colocalized throughout neuronal somata and processes as well as in astrocytes. Expression of D2 dopamine receptors in FLN-A-deficient M2 melanoma cells resulted in predominant intracellular localization of the D2 receptors, whereas in FLN-A-reconstituted cells, the D2 receptor was predominantly localized at the plasma membrane. These results suggest that FLN-A may be required for proper cell surface expression of the D2 dopamine receptors, Association of D2 and D3 dopamine receptors with FLN-A provides a mechanism whereby specific dopamine receptor subtypes may be functionally linked to downstream signaling components via the actin cytoskeleton.