SNF8, a member of the ESCRT-II complex, interacts with TRPC6 and enhances its channel activity.
SNF8, a member of the ESCRT-II complex, interacts with TRPC6 and enhances its channel activity.
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DOI：
10.1186/1471-2121-13-33
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发表时间：
2012-11-21
期刊：
影响因子：
--
通讯作者：
Schlöndorff J
中科院分区：
文献类型：
--
作者：
Carrasquillo R;Tian D;Krishna S;Pollak MR;Greka A;Schlöndorff J
瞬时受体电位经典（TRPC）通道是一种非选择性阳离子通道，参与多种细胞和组织中受体介导的钙信号传导。经典瞬时受体电位6（TRPC 6）已涉及几种病理过程，包括局灶节段性肾小球硬化症（FSGS），心脏肥大和肺动脉高压。阳离子通道的两个大的胞质区段在通道活性的适当调节中起关键作用，并且参与几种蛋白质-蛋白质相互作用。在这里，我们报告说，SNF 8，内体分选复合物转运-II（ESCRT-II）复合物的一个组成部分，与TRPC 6相互作用。这种相互作用最初是在酵母双杂交筛选中观察到的，该筛选使用TRPC 6的氨基末端胞质结构域作为诱饵，并通过真核细胞提取物的免疫共沉淀来证实。氨基末端的107个氨基酸是相互作用所必需和足够的。SNF 8的过表达增强HEK 293 T细胞中野生型和功能获得性突变体TRPC 6介导的全细胞电流此外，功能获得性突变体对NFAT介导的转录的激活通过SNF 8的过表达而增强，并且通过RNAi介导的SNF 8敲低而部分抑制。尽管ESCRT-II复合物在跨膜蛋白的内吞作用和溶酶体降解中起作用，但SNF 8过表达并不改变细胞表面上存在的TRPC 6的量。SNF 8是TRPC 6的新型结合伴侣，与通道的氨基末端胞质结构域结合。调节SNF 8表达水平改变TRPC 6通道电流，并且可以调节功能获得性突变体TRPC 6下游NFAT介导的转录的激活。总之，这些结果将SNF 8鉴定为TRPC 6的新型调节剂。
Transient receptor potential canonical (TRPC) channels are non-selective cation channels involved in receptor-mediated calcium signaling in diverse cells and tissues. The canonical transient receptor potential 6 (TRPC6) has been implicated in several pathological processes, including focal segmental glomerulosclerosis (FSGS), cardiac hypertrophy, and pulmonary hypertension. The two large cytoplasmic segments of the cation channel play a critical role in the proper regulation of channel activity, and are involved in several protein-protein interactions. Here we report that SNF8, a component of the endosomal sorting complex for transport-II (ESCRT-II) complex, interacts with TRPC6. The interaction was initially observed in a yeast two-hybrid screen using the amino-terminal cytoplasmic domain of TRPC6 as bait, and confirmed by co-immunoprecipitation from eukaryotic cell extracts. The amino-terminal 107 amino acids are necessary and sufficient for the interaction. Overexpression of SNF8 enhances both wild-type and gain-of-function mutant TRPC6-mediated whole-cell currents in HEK293T cells. Furthermore, activation of NFAT-mediated transcription by gain-of-function mutants is enhanced by overexpression of SNF8, and partially inhibited by RNAi mediated knockdown of SNF8. Although the ESCRT-II complex functions in the endocytosis and lysosomal degradation of transmembrane proteins, SNF8 overexpression does not alter the amount of TRPC6 present on the cell surface. SNF8 is novel binding partner of TRPC6, binding to the amino-terminal cytoplasmic domain of the channel. Modulating SNF8 expression levels alters the TRPC6 channel current and can modulate activation of NFAT-mediated transcription downstream of gain-of-function mutant TRPC6. Taken together, these results identify SNF8 as a novel regulator of TRPC6.
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影响因子：
3.7
作者：
Heeringa SF;Möller CC;Du J;Yue L;Hinkes B;Chernin G;Vlangos CN;Hoyer PF;Reiser J;Hildebrandt F
通讯作者：
Hildebrandt F
影响因子：
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作者：
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通讯作者：
Tiruppathi, Chinnaswamy
影响因子：
4
作者：
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通讯作者：
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影响因子：
4.8
作者：
Hisatsune, C;Kuroda, Y;Mikoshiba, K
通讯作者：
Mikoshiba, K
影响因子：
4.8
作者：
Kini, Vidisha;Chavez, Alejandra;Mehta, Dolly
通讯作者：
Mehta, Dolly