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首页> 外文期刊>Molecular and Cellular Biology >Interactions of Pex7p and Pex18p/Pex21p with the Peroxisomal Docking Machinery: Implications for the First Steps in PTS2 Protein Import
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Interactions of Pex7p and Pex18p/Pex21p with the Peroxisomal Docking Machinery: Implications for the First Steps in PTS2 Protein Import

机译：Pex7p和Pex18p / Pex21p与过氧化物酶体对接机制的相互作用：PTS2蛋白导入的第一步的含义。

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Peroxisomal PTS2-dependent matrix protein import starts with the recognition of the PTS2 targeting signal by the import receptor Pex7p. Subsequently, the formed Pex7p/cargo complex is transported from the cytosol to the peroxisomal docking complex, consisting of Pex13p and Pex14p. In Saccharomyces cerevisiae, the latter event is thought to require the redundant Pex18p and Pex21p. Here we mapped the Pex7p interaction domain of Pex13p to its N-terminal 100 amino acids. Pex18p and Pex21p also interacted with this region, albeit only in the presence of Pex7p. Expression of an N-terminally deleted version of Pex13p in a pex13Δ mutant failed to restore growth on fatty acids due to a specific defect in the import of PTS2-containing proteins. We further show by yeast two-hybrid analysis, coimmunoprecipitation, and in vitro binding assays that Pex7p can bind Pex13p and Pex14p in the absence of Pex18p/Pex21p. The PTS2 protein thiolase was shown to interact with Pex14p but not with Pex13p in a Pex7p- and Pex18p/Pex21p-dependent manner, suggesting that only Pex14p binds cargo-loaded PTS2 receptor. We also found that the cytosolic Pex7p/thiolase-containing complex includes Pex18p. This complex accumulated in docking mutants but was absent in cells lacking Pex18p/Pex21p, indicating that Pex18p/Pex21p are required already before the docking event.
机译：过氧化物酶体依赖PTS2的基质蛋白的导入始于导入受体Pex7p对PTS2靶向信号的识别。随后，将形成的Pex7p /货物复合物从胞质溶胶运输到过氧化物酶体对接复合物，该复合物由Pex13p和Pex14p组成。在酿酒酵母中，后一种事件被认为需要多余的Pex18p和Pex21p。在这里，我们将Pex13p的Pex7p相互作用域映射到其N端100个氨基酸。尽管仅在存在Pex7p的情况下，Pex18p和Pex21p仍与该区域相互作用。由于在含有PTS2的蛋白质输入中存在特定缺陷，在 pex13 Δ突变体中Nex末端缺失的Pex13p表达无法恢复脂肪酸的生长。我们进一步通过酵母双杂交分析，免疫共沉淀和体外结合试验显示，在没有Pex18p / Pex21p的情况下，Pex7p可以结合Pex13p和Pex14p。已显示PTS2蛋白硫解酶以Pex7p和Pex18p / Pex21p依赖性方式与Pex14p相互作用，但不与Pex13p相互作用，这表明只有Pex14p结合载有货物的PTS2受体。我们还发现，含胞质Pex7p /巯基酶的复合物包括Pex18p。该复合物在对接突变体中积累，但在缺少Pex18p / Pex21p的细胞中不存在，这表明在对接事件之前已经需要Pex18p / Pex21p。

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