Winther Anja Ruud, Kjos Morten, Herigstad Marie Leangen, Håvarstein Leiv Sigve, Straume Daniel
Faculty of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences, 1430 Ås, Norway.
J Bacteriol. 2021 May 1;203(9). doi: 10.1128/JB.00691-20. Epub 2021 Feb 8.
The ellipsoid shape of is determined by the synchronized actions of the elongasome and the divisome, which have the task of creating a protective layer of peptidoglycan (PG) enveloping the cell membrane. The elongasome is necessary for expanding PG in the longitudinal direction whereas the divisome synthesizes the PG that divides one cell into two. Although there is still little knowledge about how these two modes of PG synthesis are coordinated, it was recently discovered that two RNA-binding proteins called EloR and KhpA are part of a novel regulatory pathway controlling elongation in EloR and KhpA form a complex that work closely with the Ser/Thr kinase StkP to regulate cell elongation. Here, we have further explored how this regulation occur. EloR/KhpA is found at midcell, a localization fully dependent on EloR. Using a bacterial two-hybrid assay we probed EloR against several elongasome proteins and found an interaction with the lytic transglycosylase homolog MltG. By using EloR as bait in immunoprecipitation assays, MltG was pulled down confirming that they are part of the same protein complex. Fluorescent microscopy demonstrated that the Jag domain of EloR is essential for EloR's midcell localization and its interaction with MltG. Since MltG is found at midcell independent of EloR, our results suggest that MltG is responsible for recruitment of the EloR/KhpA complex to the division zone to regulate cell elongation. Bacterial cell division has been a successful target for antimicrobial agents for decades. How different pathogens regulate cell division is, however, poorly understood. To fully exploit the potential for future antibiotics targeting cell division, we need to understand the details of how the bacteria regulate and construct cell wall during this process. Here we have revealed that the newly identified EloR/KhpA complex, regulating cell elongation in , forms a complex with the essential peptidoglycan transglycosylase MltG at midcell. EloR, KhpA and MltG are conserved among many bacterial species and the EloR/KhpA/MltG regulatory pathway is most likely a common mechanism employed by many Gram-positive bacteria to coordinate cell elongation and septation.
的椭圆形形状由伸长体和分裂体的同步作用决定,它们的任务是形成一层包围细胞膜的肽聚糖(PG)保护层。伸长体对于在纵向扩展PG是必需的,而分裂体合成将一个细胞分成两个细胞的PG。尽管对于这两种PG合成模式如何协调仍知之甚少,但最近发现两种称为EloR和KhpA的RNA结合蛋白是控制中伸长的新型调控途径的一部分。EloR和KhpA形成一个复合物,与丝氨酸/苏氨酸激酶StkP密切合作以调节细胞伸长。在这里,我们进一步探讨了这种调节是如何发生的。EloR/KhpA在细胞中部被发现,其定位完全依赖于EloR。使用细菌双杂交试验,我们用几种伸长体蛋白探测EloR,发现它与溶菌转糖基酶同源物MltG相互作用。通过在免疫沉淀试验中使用EloR作为诱饵,MltG被拉下,证实它们是同一蛋白复合物的一部分。荧光显微镜显示,EloR的Jag结构域对于EloR在细胞中部的定位及其与MltG的相互作用至关重要。由于MltG在细胞中部独立于EloR被发现,我们的结果表明MltG负责将EloR/KhpA复合物招募到分裂区以调节细胞伸长。几十年来,细菌细胞分裂一直是抗菌剂的成功靶点。然而,不同病原体如何调节细胞分裂却知之甚少。为了充分利用未来靶向细胞分裂的抗生素的潜力,我们需要了解细菌在此过程中如何调节和构建细胞壁的细节。在这里,我们揭示了新鉴定的调节中细胞伸长的EloR/KhpA复合物在细胞中部与必需的肽聚糖转糖基酶MltG形成复合物。EloR、KhpA和MltG在许多细菌物种中是保守的,EloR/KhpA/MltG调控途径很可能是许多革兰氏阳性细菌用来协调细胞伸长和隔膜形成的常见机制。