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MarR 家族调控因子 OsbR 控制紫色色杆菌的氧化应激反应、厌氧硝酸盐呼吸和生物膜形成。

The MarR family regulator OsbR controls oxidative stress response, anaerobic nitrate respiration, and biofilm formation in Chromobacterium violaceum.

机构信息

Departamento de Biologia Celular e Molecular e Bioagentes Patogênicos, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP, Brazil.

Departamento de Bioquímica e Imunologia, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP, Brazil.

出版信息

BMC Microbiol. 2021 Nov 4;21(1):304. doi: 10.1186/s12866-021-02369-x.

Abstract

BACKGROUND

Chromobacterium violaceum is an environmental opportunistic pathogen that causes rare but deadly infections in humans. The transcriptional regulators that C. violaceum uses to sense and respond to environmental cues remain largely unknown.

RESULTS

Here, we described a novel transcriptional regulator in C. violaceum belonging to the MarR family that we named OsbR (oxidative stress response and biofilm formation regulator). Transcriptome profiling by DNA microarray using strains with deletion or overexpression of osbR showed that OsbR exerts a global regulatory role in C. violaceum, regulating genes involved in oxidative stress response, nitrate reduction, biofilm formation, and several metabolic pathways. EMSA assays showed that OsbR binds to the promoter regions of several OsbR-regulated genes, and the in vitro DNA binding activity was inhibited by oxidants. We demonstrated that the overexpression of osbR caused activation of ohrA even in the presence of the repressor OhrR, which resulted in improved growth under organic hydroperoxide treatment, as seem by growth curve assays. We showed that the proper regulation of the nar genes by OsbR ensures optimal growth of C. violaceum under anaerobic conditions by tuning the reduction of nitrate to nitrite. Finally, the osbR overexpressing strain showed a reduction in biofilm formation, and this phenotype correlated with the OsbR-mediated repression of two gene clusters encoding putative adhesins.

CONCLUSIONS

Together, our data indicated that OsbR is a MarR-type regulator that controls the expression of a large number of genes in C. violaceum, thereby contributing to oxidative stress defense (ohrA/ohrR), anaerobic respiration (narK1K2 and narGHJI), and biofilm formation (putative RTX adhesins).

摘要

背景

粘质沙雷氏菌是一种环境机会致病菌,可导致人类罕见但致命的感染。粘质沙雷氏菌用于感知和响应环境线索的转录调节剂在很大程度上尚不清楚。

结果

在这里,我们描述了粘质沙雷氏菌中属于 MarR 家族的一种新型转录调节剂,我们将其命名为 OsbR(氧化应激反应和生物膜形成调节剂)。使用 osbR 缺失或过表达的菌株进行 DNA 微阵列转录组分析表明,OsbR 在粘质沙雷氏菌中发挥全局调控作用,调控参与氧化应激反应、硝酸盐还原、生物膜形成和几种代谢途径的基因。EMSA 测定表明,OsbR 结合到几个 OsbR 调控基因的启动子区域,并且体外 DNA 结合活性被氧化剂抑制。我们证明,即使在抑制剂 OhrR 存在的情况下,osbR 的过表达也会导致 ohrA 的激活,从而导致在有机过氧化物处理下的生长增强,如生长曲线测定所示。我们表明,OsbR 对 nar 基因的适当调节通过调整硝酸盐还原为亚硝酸盐来确保粘质沙雷氏菌在厌氧条件下的最佳生长。最后,过表达 osbR 的菌株表现出生物膜形成减少的现象,这种表型与 OsbR 介导的两个编码假定黏附素的基因簇的抑制相关。

结论

总之,我们的数据表明 OsbR 是一种 MarR 型调节剂,可控制粘质沙雷氏菌中大量基因的表达,从而有助于氧化应激防御(ohrA/ohrR)、厌氧呼吸(narK1K2 和 narGHJI)和生物膜形成(假定的 RTX 黏附素)。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cec0/8567585/318a939f2f81/12866_2021_2369_Fig1_HTML.jpg

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