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细胞外功能(ECF)σ因子σF 参与新月柄杆菌对重金属胁迫的响应。

Extracytoplasmic function (ECF) sigma factor σF is involved in Caulobacter crescentus response to heavy metal stress.

机构信息

Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, Av, Prof, Lineu Prestes, 748, 05508-000, São Paulo, SP, Brazil.

出版信息

BMC Microbiol. 2012 Sep 18;12:210. doi: 10.1186/1471-2180-12-210.

Abstract

BACKGROUND

The α-proteobacterium Caulobacter crescentus inhabits low-nutrient environments and can tolerate certain levels of heavy metals in these sites. It has been reported that C. crescentus responds to exposure to various heavy metals by altering the expression of a large number of genes.

RESULTS

In this work, we show that the ECF sigma factor σF is one of the regulatory proteins involved in the control of the transcriptional response to chromium and cadmium. Microarray experiments indicate that σF controls eight genes during chromium stress, most of which were previously described as induced by heavy metals. Surprisingly, σF itself is not strongly auto-regulated under metal stress conditions. Interestingly, σF-dependent genes are not induced in the presence of agents that generate reactive oxygen species. Promoter analyses revealed that a conserved σF-dependent sequence is located upstream of all genes of the σF regulon. In addition, we show that the second gene in the sigF operon acts as a negative regulator of σF function, and the encoded protein has been named NrsF (Negative regulator of sigma F). Substitution of two conserved cysteine residues (C131 and C181) in NrsF affects its ability to maintain the expression of σF-dependent genes at basal levels. Furthermore, we show that σF is released into the cytoplasm during chromium stress and in cells carrying point mutations in both conserved cysteines of the protein NrsF.

CONCLUSION

A possible mechanism for induction of the σF-dependent genes by chromium and cadmium is the inactivation of the putative anti-sigma factor NrsF, leading to the release of σF to bind RNA polymerase core and drive transcription of its regulon.

摘要

背景

α变形菌新月柄杆菌栖息在营养物质较少的环境中,并能在这些环境中耐受一定水平的重金属。据报道,新月柄杆菌通过改变大量基因的表达来应对各种重金属的暴露。

结果

在这项工作中,我们表明 ECF σ 因子σF 是参与控制对铬和镉的转录反应的调节蛋白之一。微阵列实验表明,σF 在铬胁迫期间控制着 8 个基因,其中大多数基因以前被描述为被重金属诱导。令人惊讶的是,σF 本身在金属胁迫条件下并没有强烈的自我调控。有趣的是,σF 依赖的基因在产生活性氧的物质存在下不会被诱导。启动子分析表明,一个保守的 σF 依赖序列位于σF 调控基因簇的所有基因的上游。此外,我们表明,sigF 操纵子中的第二个基因作为 σF 功能的负调节剂,编码的蛋白已被命名为 NrsF(σF 的负调节剂)。NrsF 中两个保守半胱氨酸残基(C131 和 C181)的取代会影响其维持 σF 依赖基因在基础水平表达的能力。此外,我们表明,σF 在铬胁迫期间释放到细胞质中,并且在携带蛋白质 NrsF 中两个保守半胱氨酸点突变的细胞中也是如此。

结论

铬和镉诱导 σF 依赖基因的可能机制是假定的反σ 因子 NrsF 的失活,导致 σF 释放到 RNA 聚合酶核心并驱动其调控基因的转录。

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