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首页> 外文期刊>Journal of bacteriology >SwsB and SafA Are Required for CwlJ-Dependent Spore Germination in Bacillus subtilis
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SwsB and SafA Are Required for CwlJ-Dependent Spore Germination in Bacillus subtilis

机译：枯草芽孢杆菌的CWLJ依赖性孢子萌发需要SWSB和SAFA

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When Bacillus subtilis spores detect nutrients, they exit dormancy through the processes of germination and outgrowth. A key step in germination is the activation of two functionally redundant cell wall hydrolases (SleB and CwlJ) that degrade the specialized cortex peptidoglycan that surrounds the spore. How these enzymes are regulated remains poorly understood. To identify additional factors that affect their activity, we used transposon sequencing to screen for synthetic germination defects in spores lacking SleB or CwlJ. Other than the previously characterized protein YpeB, no additional factors were found to be specifically required for SleB activity. In contrast, our screen identified SafA and YlxY (renamed SwsB) in addition to the known factors GerQ and CotE as proteins required for CwlJ function. SafA is a member of the spore’s proteinaceous coat and we show that, like GerQ and CotE, it is required for accumulation and retention of CwlJ in the dormant spore. SwsB is broadly conserved among spore formers, and we show that it is required for CwlJ to efficiently degrade the cortex during germination. Intriguingly, SwsB resembles polysaccharide deacetylases, and its putative catalytic residues are required for its role in germination. However, we find no chemical signature of its activity on the spore cortex or in vitro . While the precise, mechanistic role of SwsB remains unknown, we explore and discuss potential activities. IMPORTANCE Spore formation in Bacillus subtilis has been studied for over half a century, and virtually every step in this developmental process has been characterized in molecular detail. In contrast, how spores exit dormancy remains less well understood. A key step in germination is the degradation of the specialized cell wall surrounding the spore called the cortex. Two enzymes (SleB and CwlJ) specifically target this protective layer, but how they are regulated and whether additional factors promote their activity are unknown. Here, we identified the coat protein SafA and a conserved but uncharacterized protein YlxY as additional factors required for CwlJ-dependent degradation of the cortex. Our analysis provides a more complete picture of this essential step in the exit from dormancy.
机译：当枯草芽孢杆菌孢子孢子检测营养物时，它们通过萌发和产物的过程退出休眠。发芽的一个关键步骤是激活两个功能冗余的细胞壁水解酶（SELB和CWLJ），其降解孢子围绕孢子的专用皮质肽肽。这些酶如何受到监管仍然很差。为了确定影响其活动的其他因素，我们使用转座子测序在缺乏SELB或CWLJ的孢子中的合成萌发缺陷筛选。除了先前表征的蛋白质YPEB之外，没有发现除了SELB活性所需的额外因素。相比之下，除了已知因素GERQ和COTE作为CWLJ功能所需的蛋白质外，我们的屏幕还确定了SAFA和YLXY（重命名的SWSB）。 Safa是孢子蛋白质外套的成员，我们认为，如Gerq和Cote，需要积累和保留休眠孢子中的CWLJ。 SWSB广泛保守孢子成型器，我们表明CWLJ需要在发芽期间有效降解皮质。有趣的是，SWSB类似于多糖的脱乙酰酶，其推定的催化残基需要其在发芽中的作用。然而，我们在孢子皮质或体外没有发现其活性的化学签名。虽然SWSB的确切机械作用仍然不为人知，但我们探索并讨论潜在的活动。枯草芽孢杆菌的重要性孢子形成已经过了半个世纪半个世纪，并且在该发育过程中的几乎每一步都以分子细节为特征。相比之下，孢子出口休眠如何仍然不太了解。发芽的一个关键步骤是围绕孢子的专用细胞壁的降解，称为皮质。两种酶（SELB和CWLJ）特异性靶向该保护层，但它们是如何调节的，以及促进其活动的其他因素是否未知。在这里，我们鉴定了外套蛋白质Safa和保守但无声的蛋白质YLXY作为Cortex的CWLJ依赖性降解所需的额外因素。我们的分析提供了从休眠状态的出口处的这一基本步骤的更完整的画面。

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