Bacterial Cell Biology and Physiology, Swammerdam Institute for Life Sciences, Faculty of Science, University of Amsterdam, Amsterdam, The Netherlands.
Institute of Microbiology & Infection and School of Biosciences, University of Birmingham, Edgbaston, Birmingham, UK.
Mol Microbiol. 2024 Nov;122(5):743-756. doi: 10.1111/mmi.15321. Epub 2024 Sep 30.
Escherichia coli has many periplasmic hydrolases to degrade and modify peptidoglycan (PG). However, the redundancy of eight PG endopeptidases makes it challenging to define specific roles to individual enzymes. Therefore, the cellular role of PBP7 (encoded by pbpG) is not clearly defined. In this work, we show that PBP7 localizes in the lateral cell envelope and at midcell. The C-terminal α-helix of PBP7 is crucial for midcell localization but not for its activity, which is dispensable for this localization. Additionally, midcell localization of PBP7 relies on the assembly of FtsZ up to FtsN in the divisome, and on the activity of PBP3. PBP7 was found to affect the assembly timing of FtsZ and FtsN in the divisome. The absence of PBP7 slows down the assembly of FtsN at midcell. The ΔpbpG mutant exhibited a weaker incorporation of the fluorescent D-amino acid HADA, reporting on transpeptidase activity, compared to wild-type cells. This could indicate reduced PG synthesis at the septum of the ΔpbpG strain, explaining the slower accumulation of FtsN and suggesting that endopeptidase-mediated PG cleavage may be a rate-limiting step for septal PG synthesis.
大肠杆菌拥有许多周质水解酶来降解和修饰肽聚糖(PG)。然而,八种 PG 内肽酶的冗余性使得难以确定单个酶的特定作用。因此,PBP7(由 pbpG 编码)的细胞作用尚未明确界定。在这项工作中,我们表明 PBP7 定位于细胞周质和中膜。PBP7 的 C 端α-螺旋对于中膜定位至关重要,但对于其活性并非必需,该活性对于此定位是可有可无的。此外,PBP7 的中膜定位依赖于分隔体中 FtsZ 至 FtsN 的组装,以及 PBP3 的活性。发现 PBP7 会影响分隔体中 FtsZ 和 FtsN 的组装时机。与野生型细胞相比,缺乏 PBP7 会减缓 FtsN 在中膜的组装。与野生型细胞相比,ΔpbpG 突变体在分隔体中对荧光 D-氨基酸 HADA 的掺入较弱,该氨基酸报告转肽酶活性。这可能表明 ΔpbpG 菌株的隔膜处 PG 合成减少,这解释了 FtsN 积累较慢的现象,并表明内肽酶介导的 PG 切割可能是隔膜 PG 合成的限速步骤。