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首页> 外文期刊>Journal of bacteriology >Searching for Protein-Protein Interactions within the Bacillus subtilis Spore Coat
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Searching for Protein-Protein Interactions within the Bacillus subtilis Spore Coat

机译：搜索枯草芽孢杆菌孢子外套中的蛋白质-蛋白质相互作用

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The capability of endospores of Bacillus subtilis to withstand extreme environmental conditions is secured by several attributes. One of them, the protein shell that encases the spore and is known as the coat, provides the spore with its characteristic resistance to toxic chemicals, lytic enzymes, and predation by unicellular and multicellular eukaryotes. Despite most of the components of the spore coat having been identified, we have only a vague understanding of how such a complex structure is assembled. Using the yeast two-hybrid system, we attempted to identify direct contacts among the proteins allocated to the insoluble fraction of the spore coat: CotV, CotW, CotX, CotY, and CotZ. We also examined whether they could interact with CotE, one of the most crucial morphogenetic proteins governing outer coat formation and also present in the insoluble fraction. Out of all 21 possible interactions we tested, 4 were found to be positive. Among these interactions, we confirmed the previous observation that CotE forms homo-oligomers. In addition, we observed homotypic interactions of CotY, strong interactions between CotZ and CotY, and relatively weak, yet significant, interactions between CotV and CotW. The results of this yeast two-hybrid analysis were confirmed by size exclusion chromatography of recombinant coat proteins and a pull-down assay.
机译：枯草芽孢杆菌的内生孢子承受极端环境条件的能力有几个属性。其中之一是包裹孢子的蛋白质外壳，被称为外壳。它为孢子提供了对有毒化学物质，裂解酶以及单细胞和多细胞真核生物的捕食的特有抗性。尽管已鉴定出孢子衣的大部分成分，但对于这种复杂结构的组装方式我们还是只有模糊的认识。使用酵母双杂交系统，我们尝试确定分配给孢子外壳的不溶部分的蛋白质之间的直接接触：CotV，CotW，CotX，CotY和CotZ。我们还检查了它们是否可以与CotE相互作用，CotE是控制外壳形成的最关键的形态发生蛋白之一，并且也存在于不溶级分中。在我们测试的所有21种可能的互动中，有4种是阳性。在这些相互作用中，我们证实了先前的观察结果，即CotE形成同源寡聚体。此外，我们观察到CotY的同型相互作用，CotZ和CotY之间的强相互作用以及CotV和CotW之间的相对弱但显着的相互作用。通过重组外壳蛋白的尺寸排阻色谱法和下拉测定法证实了该酵母两杂交分析的结果。

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