Drosophila Vps35 function is necessary for normal endocytic trafficking and actin cytoskeleton organisation
Drosophila Vps35 function is necessary for normal endocytic trafficking and actin cytoskeleton organisation
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DOI：
10.1242/jcs.012336
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发表时间：
2007-12-15
影响因子：
4
通讯作者：
O'Kane, Cahir J.
中科院分区：
文献类型：
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作者：
Korolchuk, Viktor I.;Schuetz, Martin M.;O'Kane, Cahir J.
为了鉴定受体介导的内吞作用所需的新蛋白质，我们在果蝇 S2 细胞中开发了一种基于 RNAi 的筛选方法，该方法基于清道夫受体配体的摄取。一些已知的内吞蛋白对于该测定中的内吞作用至关重要，包括网格蛋白和α-适应素；然而，不需要对突触小泡内吞作用重要的其他蛋白质。在对新型内吞蛋白的小规模筛选中，我们鉴定了 Vps35 的果蝇同源物，它是逆转录体复合物的一个组成部分，参与内体到高尔基体的运输。 Vps35 的缺失会抑制清道夫受体配体的内吞作用，并导致许多受体和内吞蛋白的错误定位。 Vps35 具有肿瘤抑制特性，因为它的缺失会导致幼虫中血细胞过度增殖。它的缺失还会导致神经肌肉接头处的信号传导缺陷，包括 TGF beta/BMP 信号传导的上调和突触末端的过度形成。 Vps35 负向调节肌动蛋白聚合，遗传相互作用表明 vps35 突变体的一些内吞和信号传导缺陷是由于这种功能造成的。
To identify novel proteins required for receptor-mediated endocytosis, we have developed an RNAi-based screening method in Drosophila S2 cells, based on uptake of a scavenger receptor ligand. Some known endocytic proteins are essential for endocytosis in this assay, including clathrin and alpha-adaptin; however, other proteins important for synaptic vesicle endocytosis are not required. In a small screen for novel endocytic proteins, we identified the Drosophila homologue of Vps35, a component of the retromer complex, involved in endosome-to-Golgi trafficking. Loss of Vps35 inhibits scavenger receptor ligand endocytosis, and causes mislocalisation of a number of receptors and endocytic proteins. Vps35 has tumour suppressor properties because its loss leads to overproliferation of blood cells in larvae. Its loss also causes signalling defects at the neuromuscular junction, including upregulation of TGF beta/BMP signalling and excessive formation of synaptic terminals. Vps35 negatively regulates actin polymerisation, and genetic interactions suggest that some of the endocytic and signalling defects of vps35 mutants are due to this function.