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细胞壁抑制反应蛋白 CwrA 在. 的致病性中的作用。

The roles of cell wall inhibition responsive protein CwrA in the pathogenicity of .

机构信息

Department of Clinical Laboratory, Shanghai Pulmonary Hospital, School of Medicine, Tongji University, Shanghai, People's Republic of China.

Shanghai Institute of Immunity and Infection, Chinese Academy of Science, Shanghai, People's Republic of China.

出版信息

Virulence. 2024 Dec;15(1):2411540. doi: 10.1080/21505594.2024.2411540. Epub 2024 Oct 5.

Abstract

The ability to form robust biofilms and secrete a diverse array of virulence factors are key pathogenic determinants of , causing a wide range of infectious diseases. Here, we characterized as a VraR-regulated gene encoding a cell wall inhibition-responsive protein (CwrA) using electrophoretic mobility shift assays. We constructed deletion mutants in the genetic background of methicillin-resistant (MRSA) and methicillin-sensitive (MSSA) strains. Phenotypic analyses indicated that deletion of led to impaired biofilm formation, which was correlated with polysaccharide intercellular adhesin (PIA). Besides, the results of real-time quantitative PCR (RT-qPCR) and β-galactosidase activity assay revealed that CwrA promoted biofilm formation by influence the operon activity in . Furthermore, deletion mutants released less extracellular DNA (eDNA) in the biofilm because of their reduced autolytic activity compared to the wild-type (WT) strains. We also found that deletion mutant more virulence than the parental strain because of its enhanced hemolytic activity. Mechanistically, this phenotypic alteration is related to activation of the SaeRS two-component system, which positively regulates the transcriptional levels of genes encoding membrane-damaging toxins. Overall, our results suggest that CwrA plays an important role in modulating biofilm formation and hemolytic activity in .

摘要

形成强大生物膜并分泌多种毒力因子的能力是 致病性的关键决定因素,可导致广泛的感染性疾病。在这里,我们使用电泳迁移率变动分析,将 鉴定为 VraR 调控的基因,该基因编码一种细胞壁抑制反应蛋白(CwrA)。我们在耐甲氧西林金黄色葡萄球菌(MRSA)和甲氧西林敏感金黄色葡萄球菌(MSSA)菌株的遗传背景下构建了 缺失突变体。表型分析表明, 缺失导致生物膜形成受损,这与多糖细胞间黏附素(PIA)有关。此外,实时定量 PCR(RT-qPCR)和β-半乳糖苷酶活性测定的结果表明,CwrA 通过影响 操纵子活性来促进生物膜的形成。此外,与野生型(WT)菌株相比, 缺失突变体在生物膜中释放的细胞外 DNA(eDNA)较少,因为其自溶活性降低。我们还发现, 缺失突变体比亲本菌株具有更高的毒力,因为其溶血活性增强。从机制上讲,这种表型改变与 SaeRS 双组分系统的激活有关,该系统正向调节编码膜损伤毒素的基因的转录水平。总的来说,我们的研究结果表明,CwrA 在调节生物膜形成和 溶血活性方面发挥着重要作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ff3a/11457683/0998270cce14/KVIR_A_2411540_F0001_OC.jpg

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