The WalRK Two-Component System Is Essential for Proper Cell Envelope Biogenesis in Clostridioides difficile.
The WalRK Two-Component System Is Essential for Proper Cell Envelope Biogenesis in Clostridioides difficile.
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DOI：
10.1128/jb.00121-22
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发表时间：
2022-06-21
影响因子：
3.2
通讯作者：
中科院分区：
文献类型：
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作者：
WalR-WalK双组分调节系统（TCS）存在于所有厚壁菌门中，其中它调节在生长和分裂期间重塑细胞包膜所需的多个基因的表达。与大多数TCS不同，WalRK对生存力至关重要，因此它作为潜在的抗生素靶标引起了人们的兴趣。在这项研究中，我们使用WalR的过表达和CRISPR干扰来研究艰难梭菌的Wal系统，这是高收入国家医院相关性腹泻的主要原因。我们证实了wal操纵子是必需的，并确定了形态缺陷和细胞溶解作为改变wal表达的主要终端表型。我们还使用转录组测序（RNA-seq）来鉴定超过150个基因，其表达响应于WalR水平而变化。该基因集富含细胞包膜基因，包括编码几种预测的PG水解酶和可以调节PG水解酶活性的蛋白质的基因。C.艰难梭菌细胞膜的一个重要特征是存在S层，我们发现WalR影响编码S层蛋白的几个基因的表达。一个意想不到的发现是，与其他硬壁菌门报道的情况相比，一些沃尔相关的表型缺陷被逆转。例如，下调Wal信号转导导致C.艰难的细胞变得更长而不是更短，如枯草芽孢杆菌。同样，下调Wal使C.艰难梭菌对万古霉素更敏感，而Wal活性降低与金黄色葡萄球菌中万古霉素耐药性增加有关。WalRK双组分系统（TCS）是厚壁菌门中协调肽聚糖合成和周转的关键。我们研究了WalRK TCS在艰难梭菌中的作用，艰难梭菌是一种具有非典型细胞包膜的重要细菌病原体。我们证实，WalRK是必不可少的，并调节细胞被膜生物合成，虽然我们观察到的几个表型变化是相反的其他厚壁菌门已报道。我们还确定了超过150个基因，其表达直接或间接由WalR控制。总的来说，我们的研究结果提供了一个重要的调控系统和潜在的抗生素靶标在C的未来调查的基础。很难
The WalR-WalK two-component regulatory system (TCS) is found in all Firmicutes, in which it regulates the expression of multiple genes required for remodeling the cell envelope during growth and division. Unlike most TCSs, WalRK is essential for viability, so it has attracted interest as a potential antibiotic target. In this study, we used overexpression of WalR and CRISPR interference to investigate the Wal system of Clostridioides difficile, a major cause of hospital-associated diarrhea in high-income countries. We confirmed that the wal operon is essential and identified morphological defects and cell lysis as the major terminal phenotypes of altered wal expression. We also used transcriptome sequencing (RNA-seq) to identify over 150 genes whose expression changes in response to WalR levels. This gene set is enriched in cell envelope genes and includes genes encoding several predicted PG hydrolases and proteins that could regulate PG hydrolase activity. A distinct feature of the C. difficile cell envelope is the presence of an S-layer, and we found that WalR affects expression of several genes which encode S-layer proteins. An unexpected finding was that some Wal-associated phenotypic defects were inverted in comparison to what has been reported for other Firmicutes. For example, downregulation of Wal signaling caused C. difficile cells to become longer rather than shorter, as in Bacillus subtilis. Likewise, downregulation of Wal rendered C. difficile more sensitive to vancomycin, whereas reduced Wal activity is linked to increased vancomycin resistance in Staphylococcus aureus. IMPORTANCE The WalRK two-component system (TCS) is essential for coordinating synthesis and turnover of peptidoglycan in Firmicutes. We investigated the WalRK TCS in Clostridioides difficile, an important bacterial pathogen with an atypical cell envelope. We confirmed that WalRK is essential and regulates cell envelope biogenesis, although several of the phenotypic changes we observed were opposite to what has been reported for other Firmicutes. We also identified over 150 genes whose expression is controlled either directly or indirectly by WalR. Overall, our findings provide a foundation for future investigations of an important regulatory system and potential antibiotic target in C. difficile.
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影响因子：
3.2
作者：
Dobihal, Genevieve S.;Flores-Kim, Josue;Rudner, David Z.
通讯作者：
Rudner, David Z.
影响因子：
14.9
作者：
Caspi R;Billington R;Ferrer L;Foerster H;Fulcher CA;Keseler IM;Kothari A;Krummenacker M;Latendresse M;Mueller LA;Ong Q;Paley S;Subhraveti P;Weaver DS;Karp PD
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Karp PD
影响因子：
3.6
作者：
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通讯作者：
Msadek, Tarek
影响因子：
2.9
作者：
Eveland, SS;Pompliano, DL;Anderson, MS
通讯作者：
Anderson, MS
影响因子：
3.6
作者：
Bisicchia, Paola;Noone, David;Devine, Kevin M.
通讯作者：
Devine, Kevin M.