FtsEX-independent control of RipA-mediated cell separation in Corynebacteriales.
FtsEX-independent control of RipA-mediated cell separation in Corynebacteriales.
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DOI：
10.1073/pnas.2214599119
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发表时间：
2022-12-13
影响因子：
11.1
通讯作者：
Alzari, Pedro M.
中科院分区：
文献类型：
--
作者：
Gaday, Quentin;Megrian, Daniela;Carloni, Giacomo;Martinez, Mariano;Sokolova, Bohdana;Ben Assaya, Mathilde;Legrand, Pierre;Brule, Sebastien;Haouz, Ahmed;Wehenkel, Anne Marie;Alzari, Pedro M.
细胞壁的合成、维持和降解是细菌生存的基本过程，也是药物开发的重要靶点。负责肽聚糖水解的酶（称为鼠蛋白水解酶）必须严格调节以避免自溶。在包括重要病原体结核分枝杆菌和白喉棒状杆菌在内的棒状杆菌目中，用于细胞分离的最重要的小鼠水解酶是内肽酶RipA。我们在此展示了来自谷氨酸棒状杆菌的全长RipA同源物Cg1735的晶体结构，它揭示了一种不寻常的自抑制模式。我们描述了这种自抑制是如何被保守的跨膜蛋白Cg1604缓解的，并提出了一个不依赖ftsex的模型，通过Cg1604 - cg1735复合物来控制PG水解。细菌细胞壁是一个多层网状结构，其主要成分是肽聚糖（PG），一种由短肽茎交联的糖聚合物。在细胞分裂过程中，PG合成和降解的谨慎平衡，在时间和空间上精确协调，是防止细胞壁失控破坏所必需的。在棒状杆菌中，D，L内肽酶RipA已成为细胞分离的主要PG水解酶，RipA缺失对人类病原体结核分枝杆菌和白喉棒状杆菌的毒力有重要影响。然而，RipA介导细胞分离的确切机制仍然难以捉摸。在这里，我们报道了RipA Cg1735的谷氨酸棒状杆菌同源物的系统发育、生化和结构分析。两种不同晶体形式的全长Cg1735的晶体结构揭示了c端NlpC/P60催化结构域被其n端保守的线圈结构域挤压，使酶处于自抑制状态。我们发现跨膜间隔蛋白Cg1604的胞外核心结构域减轻了这种自抑制作用。Cg1604的晶体结构显示了一个（β/α）蛋白，其整体拓扑结构与响应调节蛋白的受体结构域相似。基于生物信息学和突变分析的Cg1735 - Cg1604复合物的原子模型表明，Cg1604中保守的远膜螺旋插入是Cg1735活化的原因。报道的数据提供了重要的见解，如何胞内细胞分裂信号(s)，尚未确定，在ripa介导的棒状杆菌细胞分离过程中，控制PG水解。
Cell wall synthesis, maintenance, and degradation are essential processes for bacteria and important targets for drug development. The enzymes responsible for peptidoglycan hydrolysis (termed murein hydrolases) must be tightly regulated to avoid autolysis. In the order Corynebacteriales, which includes the important pathogens Mycobacterium tuberculosis and Corynebacterium diphtheriae, the most important murein hydrolase for cell separation is the endopeptidase RipA. We present here the crystal structure of a full-length RipA homologue from Corynebacterium glutamicum, Cg1735, which reveals an unusual mode of autoinhibition. We describe how this autoinhibition is relieved by the conserved transmembrane protein Cg1604 and propose an FtsEX-independent model for septal control of PG hydrolysis via the Cg1604–Cg1735 complex. The bacterial cell wall is a multi-layered mesh, whose major component is peptidoglycan (PG), a sugar polymer cross-linked by short peptide stems. During cell division, a careful balance of PG synthesis and degradation, precisely coordinated both in time and space, is necessary to prevent uncontrolled destruction of the cell wall. In Corynebacteriales, the D,L endopeptidase RipA has emerged as a major PG hydrolase for cell separation, and RipA defaults have major implications for virulence of the human pathogens Mycobacterium tuberculosis and Corynebacterium diphtheriae. However, the precise mechanisms by which RipA mediates cell separation remain elusive. Here we report phylogenetic, biochemical, and structural analysis of the Corynebacterium glutamicum homologue of RipA, Cg1735. The crystal structures of full-length Cg1735 in two different crystal forms revealed the C-terminal NlpC/P60 catalytic domain obtruded by its N-terminal conserved coiled-coil domain, which locks the enzyme in an autoinhibited state. We show that this autoinhibition is relieved by the extracellular core domain of the transmembrane septal protein Cg1604. The crystal structure of Cg1604 revealed a (β/α) protein with an overall topology similar to that of receiver domains from response regulator proteins. The atomic model of the Cg1735–Cg1604 complex, based on bioinformatical and mutational analysis, indicates that a conserved, distal-membrane helical insertion in Cg1604 is responsible for Cg1735 activation. The reported data provide important insights into how intracellular cell division signal(s), yet to be identified, control PG hydrolysis during RipA-mediated cell separation in Corynebacteriales.
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影响因子：
64.8
作者：
Jumper J;Evans R;Pritzel A;Green T;Figurnov M;Ronneberger O;Tunyasuvunakool K;Bates R;Žídek A;Potapenko A;Bridgland A;Meyer C;Kohl SAA;Ballard AJ;Cowie A;Romera-Paredes B;Nikolov S;Jain R;Adler J;Back T;Petersen S;Reiman D;Clancy E;Zielinski M;Steinegger M;Pacholska M;Berghammer T;Bodenstein S;Silver D;Vinyals O;Senior AW;Kavukcuoglu K;Kohli P;Hassabis D
通讯作者：
Hassabis D
影响因子：
6.4
作者：
DeJesus MA;Gerrick ER;Xu W;Park SW;Long JE;Boutte CC;Rubin EJ;Schnappinger D;Ehrt S;Fortune SM;Sassetti CM;Ioerger TR
通讯作者：
Ioerger TR
影响因子：
6.7
作者：
Chao, Michael C.;Kieser, Karen J.;Rubin, Eric J.
通讯作者：
Rubin, Eric J.
影响因子：
64.5
作者：
Fiumara F;Fioriti L;Kandel ER;Hendrickson WA
通讯作者：
Hendrickson WA
DOI：
10.1093/bioinformatics/btp348
发表时间：
2009-08-01
期刊：
Bioinformatics (Oxford, England)
影响因子：
--
作者：
Capella-Gutiérrez S;Silla-Martínez JM;Gabaldón T
通讯作者：
Gabaldón T