Proteins Involved in Formation of the Outermost Layer of Bacillus subtilis Spores
Proteins Involved in Formation of the Outermost Layer of Bacillus subtilis Spores
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DOI：
10.1128/jb.05310-11
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发表时间：
2011-08-01
影响因子：
3.2
通讯作者：
Watabe, Kazuhito
中科院分区：
文献类型：
--
作者：
Imamura, Daisuke;Kuwana, Ritsuko;Watabe, Kazuhito
为了研究枯草芽孢杆菌孢子的最外层结构，我们分析了针对枯草杆菌孢子上蛋白质的抗体的可及性。抗绿色荧光蛋白(GFP)抗体有效地获得了与CgeA或Cotz融合的GFP，这些GFP以前被分配到最外面的一层，称为孢子壳。然而，抗GFP抗体不能与融合了14种外衣、内衣或皮质蛋白的GFP表达菌株的孢子结合。抗CgeA抗体可与野生型和CgeA-GFP菌株的孢子结合，但不能与CgeA突变菌株的孢子结合。这些结果表明，孢子壳覆盖着孢子被层，是枯草杆菌孢子最外层的外露部分。我们发现，Cotz是孢子壳包围孢子所必需的，但不是形成孢子皮所必需的，这表明Cotz在孢子皮的形成中起着关键作用。此外，我们还发现Coty-GFP暴露在孢子表面，表明Coty是孢子壳的一个附加成分。此外，Coty-GFP在孢子周围的定位依赖于Cotz，而Coty和Cotz相互依赖于孢子的组装。此外，CoTW的破坏影响了CotV-GFP的组装，而CotX的破坏影响了CotV-GFP和CgeA-GFP的组装。这些结果表明，coVWXYZ簇中的cgeA和基因参与了孢子壳的形成。
To investigate the outermost structure of the Bacillus subtilis spore, we analyzed the accessibility of antibodies to proteins on spores of B. subtilis. Anti-green fluorescent protein (GFP) antibodies efficiently accessed GFP fused to CgeA or CotZ, which were previously assigned to the outermost layer termed the spore crust. However, anti-GFP antibodies did not bind to spores of strains expressing GFP fused to 14 outer coat, inner coat, or cortex proteins. Anti-CgeA antibodies bound to spores of wild-type and CgeA-GFP strains but not cgeA mutant spores. These results suggest that the spore crust covers the spore coat and is the externally exposed, outermost layer of the B. subtilis spore. We found that CotZ was essential for the spore crust to surround the spore but not for spore coat formation, indicating that CotZ plays a critical role in spore crust formation. In addition, we found that CotY-GFP was exposed on the surface of the spore, suggesting that CotY is an additional component of the spore crust. Moreover, the localization of CotY-GFP around the spore depended on CotZ, and CotY and CotZ depended on each other for spore assembly. Furthermore, a disruption of cotW affected the assembly of CotV-GFP, and a disruption of cotX affected the assembly of both CotV-GFP and CgeA-GFP. These results suggest that cgeA and genes in the cotVWXYZ cluster are involved in spore crust formation.