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首页> 外文期刊>Molecular biology of the cell >PIP promotes the recruitment of the GGA adaptor proteins to the trans-Golgi TS recognition of the ubiquitin network and regulates their sorting signal
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PIP promotes the recruitment of the GGA adaptor proteins to the trans-Golgi TS recognition of the ubiquitin network and regulates their sorting signal

机译：PIP促进GGA衔接子蛋白的募集，以泛素网络的反式高尔基TS识别并调节其分选信号

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摘要

Phosphatidylinositol 4 phosphate (PI4P) is highly enriched in the trans-Golgi network (TGN). Here we establish that PI4P is a key regulator of the recruitment of the GGA clathrin adaptor proteins to the TGN and that PI4P has a novel role in promoting their recognition of the ubiquitin (Ub) sorting signal. Knockdown of PI4KII alpha by RNA interference (RNAi), which depletes the TGN's PI4P, impaired the recruitment of the GGAs to the TGN. GGAs bind PI4P primarily through their GAT domain, in a region called C-GAT, which also binds Ub but not Arf1. We identified two basic residues in the GAT domain that are essential for PI4P binding in vitro and for the recruitment of GGAs to the TGN in vivo. Unlike wild-type GGA, GGA with mutated GATs failed to rescue the abnormal TGN phenotype of the GGA RNAi-depleted cells. These residues partially overlap with those that bind Ub, and PI4P increased the affinity of the GAT domain for Ub. Because the recruitment of clathrin adaptors and their cargoes to the TGN is mediated through a web of low-affinity interactions, our results show that the dual roles of PI4P can promote specific GGA targeting and cargo recognition at the TGN.
机译：磷酸磷脂酰肌醇4磷酸（PI4P）在反高尔基网络（TGN）中高度富集。在这里，我们确定PI4P是GGA网格蛋白衔接子蛋白向TGN募集的关键调节剂，并且PI4P在促进其对泛素（Ub）分类信号的识别中具有新作用。 RNA干扰（RNAi）抑制了PI4KII alpha，这耗尽了TGN的PI4P，从而损害了GGA向TGN的募集。 GGA主要通过其GAT域（称为C-GAT）结合PI4P，该区域也结合Ub但不结合Arf1。我们在GAT域中确定了两个基本残基，这些残基对于体外PI4P结合以及体内GGA募集到TGN是必不可少的。与野生型GGA不同，具有突变GAT的GGA无法挽救GGA RNAi耗尽细胞的异常TGN表型。这些残基与结合Ub的残基部分重叠，并且PI4P增加了GAT结构域对Ub的亲和力。由于网格蛋白衔接子及其货物的募集是通过低亲和力相互作用网络介导的，因此我们的结果表明PI4P的双重作用可以促进TGA上特定的GGA靶向和货物识别。

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