A new D,L-Endopeptidase gene product, YojL (renamed CwlS), plays a role in cell separation with LytE and LytF in Bacillus subtilis
A new D,L-Endopeptidase gene product, YojL (renamed CwlS), plays a role in cell separation with LytE and LytF in Bacillus subtilis
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DOI：
10.1128/jb.00188-06
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发表时间：
2006-08-01
影响因子：
3.2
通讯作者：
Sekiguchi, Junichi
中科院分区：
文献类型：
--
作者：
Fukushima, Tatsuya;Afkham, Anahita;Sekiguchi, Junichi
一种新的肽聚糖水解酶枯草芽孢杆菌YojL（与细胞分离相关的细胞壁溶解酶，更名为CwlS）与与细胞分离相关的LytE（CwlF）和LytF（CwlE）具有高度的氨基酸序列相似性。 CwlS 的 N 末端区域有四个串联重复区域（LysM 重复区域），预计是肽聚糖结合模块。 C 末端区域与 LytE 和 LytF 的 C 末端细胞壁水解酶结构域表现出高度相似性。大肠杆菌产生的CwlS的C端区域可以水解枯草芽孢杆菌细胞壁中的D-γ-谷氨酰-内消旋-二氨基-庚二酸键，表明CwlS是一种D,L-内肽酶。 β-半乳糖苷酶融合实验和 Northern 杂交分析表明 cwlS 基因在营养晚期和稳定早期转录。 cwlS突变体表现出与野生型相似的细胞形状；然而，lytE lytF cwlS 三重突变体表现出聚集的微纤维形成。此外，免疫荧光显微镜显示，FLAG标记的CwlS位于营养后期的细胞分离位点和细胞极。定位位点与 LytF 和 LytE 相似，表明 CwlS 参与了 LytF 和 LytE 的细胞分离。这些特定的定位可能取决于其 N 端结构域中的 LysM 重复序列。讨论了 CwlS、LytF 和 LytE 在细胞分离中的作用。
A new peptidoglycan hydrollase, Bacillus subtilis YojL (cell wall-lytic enzyme associated with cell separation, renamed CwlS), exhibits high amino acid sequence similarity to LytE (CwlF) and LytF (CwlE), which are associated with cell separation. The N-terminal region of CwlS has four tandem repeat regions (LysM repeats) predicted to be a peptidoglycan-binding module. The C-terminal region exhibits high similarity to the cell wall hydrolase domains of LytE and LytF at their C-terminal ends. The C-terminal region of CwlS produced in Escherichia coli could hydrolyze the linkage of D-gamma-glutamyl-meso-diamino-pimelic acid in the cell wall of B. subtilis, suggesting that CwlS is a D,L-endopeptidase. beta-Galactosidase fusion experiments and Northern hybridization analysis suggested that the cwlS gene is transcribed during the late vegetative and early stationary phases. A cwlS mutant exhibited a cell shape similar to that of the wild type; however, a lytE lytF cwlS triple mutant exhibited aggregated microfiber formation. Moreover, immunofluorescence microscopy showed that FLAG-tagged CwlS was localized at cell separation sites and cell poles during the late vegetative phase. The localization sites are similar to those of LytF and LytE, indicating that CwlS is involved in cell separation with LytF and LytE. These specific localizations may be dependent on the LysM repeats in their N-terminal domains. The roles of CwlS, LytF, and LytE in cell separation are discussed.