A cortex-specific penicillin-binding protein contributes to heat resistance in Clostridioides difficile spores.
A cortex-specific penicillin-binding protein contributes to heat resistance in Clostridioides difficile spores.
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DOI：
10.1016/j.anaerobe.2021.102379
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发表时间：
2021-08
期刊：
影响因子：
2.3
通讯作者：
Fagan RP
中科院分区：
文献类型：
--
作者：
Alabdali YAJ;Oatley P;Kirk JA;Fagan RP
孢子形成是厚壁菌门的许多成员共有的复杂细胞分化程序，其最终结果是可以在恶劣的环境损伤中存活的高度抗性、代谢惰性的孢子。艰难梭菌孢子是传播疾病所必需的，也是反复感染所必需的。然而，孢子形成的分子基础知之甚少，尽管与充分研究的枯草芽孢杆菌系统相似。孢子被膜由多个保护层组成，其中一层是称为皮层的肽聚糖的专门层，这对孢子的抗性特性至关重要。我们着手鉴定艰难梭菌中合成皮质肽聚糖所需的酶。对艰难梭菌基因组进行生物信息学分析以鉴定枯草芽孢杆菌spoVD的推定同源物，与定向诱变和显微镜检查相结合以鉴定表皮特异性PBP活性。CDR20291_2544（SpoVDCd）的缺失消除了孢子形成，并且通过顺式互补完全恢复了该表型。SpoVDCd的分析揭示了由二聚化、转肽酶和PASTA结构域组成的三结构域结构，非常类似于B. subtilis SpoVD。与SpoVDCd结构域突变体的互补表明PASTA结构域是形成形态正常的孢子所必需的。SpoVDCd也被认为是本地化的发展孢子超分辨率共聚焦显微镜。我们已经鉴定并表征了艰难梭菌中的皮质特异性PBP。这是艰难梭菌中皮质特异性PBP的第一个表征，并开始解开这种重要病原体中皮质生物发生的过程。CDR 20291_2544编码枯草B SpoVD的艰难梭菌同源物。spoVDCd的突变完全阻止了耐热孢子的形成。SpoVDCd PASTA结构域对于其功能来说是可有可无的。SpoVDCd定位于发育中的孢子。
Sporulation is a complex cell differentiation programme shared by many members of the Firmicutes, the end result of which is a highly resistant, metabolically inert spore that can survive harsh environmental insults. Clostridioides difficile spores are essential for transmission of disease and are also required for recurrent infection. However, the molecular basis of sporulation is poorly understood, despite parallels with the well-studied Bacillus subtilis system. The spore envelope consists of multiple protective layers, one of which is a specialised layer of peptidoglycan, called the cortex, that is essential for the resistant properties of the spore. We set out to identify the enzymes required for synthesis of cortex peptidoglycan in C. difficile. Bioinformatic analysis of the C. difficile genome to identify putative homologues of Bacillus subtilis spoVD was combined with directed mutagenesis and microscopy to identify and characterise cortex-specific PBP activity. Deletion of CDR20291_2544 (SpoVDCd) abrogated spore formation and this phenotype was completely restored by complementation in cis. Analysis of SpoVDCd revealed a three domain structure, consisting of dimerization, transpeptidase and PASTA domains, very similar to B. subtilis SpoVD. Complementation with SpoVDCd domain mutants demonstrated that the PASTA domain was dispensable for formation of morphologically normal spores. SpoVDCd was also seen to localise to the developing spore by super-resolution confocal microscopy. We have identified and characterised a cortex specific PBP in C. difficile. This is the first characterisation of a cortex-specific PBP in C. difficile and begins the process of unravelling cortex biogenesis in this important pathogen. CDR20291_2544 encodes a C. difficile homologue of the B subtilis SpoVD. Mutation of spoVDCd completely prevents the formation of heat-resistant spores. The SpoVDCd PASTA domain was dispensable for its function. SpoVDCd localises to the developing spore.
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影响因子：
2.3
作者：
Kirk, Joseph A.;Fagan, Robert P.
通讯作者：
Fagan, Robert P.
影响因子：
3.7
作者：
Ng YK;Ehsaan M;Philip S;Collery MM;Janoir C;Collignon A;Cartman ST;Minton NP
通讯作者：
Minton NP
影响因子：
4.8
作者：
Fagan, Robert P.;Fairweather, Neil F.
通讯作者：
Fairweather, Neil F.
影响因子：
4.8
作者：
Peltier, Johann;Courtin, Pascal;Pons, Jean-Louis
通讯作者：
Pons, Jean-Louis
影响因子：
3.1
作者：
Deakin, Laura J.;Clare, Simon;Lawley, Trevor D.
通讯作者：
Lawley, Trevor D.