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Deletion mutants of AP-1 adaptin subunits display distinct phenotypes in fission yeast

机译：AP-1适应蛋白亚基的缺失突变体在裂殖酵母中表现出不同的表型

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摘要

Adaptins are subunits of the heterotetrameric (o/o/d/s) adaptor protein (AP) complexes that are involved in clathrin-mediated membrane trafficking. Here, we show that in Schizosaccharomyces pombe the deletion strains of each individual subunit of the AP-1 complex [Apl2 (o), Apl4 (d), Apm1 (o) and Aps1 (s)] caused distinct phenotypes on growth sensitivity to temperature or drugs. We also show that the apm1 and apl2 mutants displayed similar but more severe phenotypes than those of aps1 or apl4 mutants. Furthermore, the apl2aps1 and apl2apl4 double mutants displayed synthetic growth defects, whereas the aps1apl4 and apl2apm1 double mutants did not. In pull-down assay, Apm1 binds Apl2 even in the absence of Aps1 and Apl4, and Apl4 binds Aps1 even in the absence of Apm1 and Apl2. Consistently, the deletion of any subunit generally caused the disassociation of the heterotetrameric complex from endosomes, although some subunits weakly localized to endosomes. In addition, the deletion of individual subunits caused similar endosomal accumulation of v-SNARE synaptobrevin Syb1. Altogether, results suggest that the four subunits are all essential for the heterotetrameric complex formation and for the AP-1 function in exit transport from endosomes.
机译：Adaptin是异四聚体（o / o / d / s）衔接蛋白（AP）复合物的亚基，参与网格蛋白介导的膜运输。在这里，我们显示在粟酒裂殖酵母中，AP-1复合物的每个单独亚基的缺失菌株[Apl2（o），Apl4（d），Apm1（o）和Aps1（s）]对生长对温度的敏感性引起了明显的表型。或毒品。我们还显示，与aps1或apl4突变体相比，apm1和apl2突变体显示相似但更严重的表型。此外，apl2aps1和apl2apl4双突变体显示出合成生长缺陷，而aps1apl4和apl2apm1双突变体则没有。在下拉测定中，即使在不存在Aps1和Apl4的情况下，Apm1也结合Apl2，并且即使在不存在Apm1和Apl2的情况下，Apl4也结合Aps1。一致地，任何亚基的缺失通常引起异四聚体复合体与内体的分离，尽管一些亚基弱地定位于内体。此外，单个亚基的缺失导致类似的内体积累的v-SNARE突触短纤维Syb1。总而言之，结果表明，这四个亚基对于异四聚体复合物的形成以及AP-1从内体的出口转运功能都是必不可少的。

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