A PadR family transcriptional repressor regulates the transcription of chromate efflux transporter in Enterobacter sp. Z1

IF 3.3 4区 生物学 Q2 MICROBIOLOGY
Xueqi Huo, Zijie Zhou, Hongliang Liu, Gejiao Wang, Kaixiang Shi
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引用次数: 0

Abstract

Chromium is a prevalent toxic heavy metal, and chromate [Cr(VI)] exhibits high mutagenicity and carcinogenicity. The presence of the Cr(VI) efflux protein ChrA has been identified in strains exhibiting resistance to Cr(VI). Nevertheless, certain strains of bacteria that are resistant to Cr(VI) lack the presence of ChrB, a known regulatory factor. Here, a PadR family transcriptional repressor, ChrN, has been identified as a regulator in the response of Enterobacter sp. Z1(CCTCC NO: M 2019147) to Cr(VI). The chrN gene is cotranscribed with the chrA gene, and the transcriptional expression of this operon is induced by Cr(VI). The binding capacity of the ChrN protein to Cr(VI) was demonstrated by both the tryptophan fluorescence assay and Ni-NTA purification assay. The interaction between ChrN and the chrAN operon promoter was validated by reporter gene assay and electrophoretic mobility shift assay. Mutation of the conserved histidine residues His14 and His50 resulted in loss of ChrN binding with the promoter of the chrAN operon. This observation implies that these residues are crucial for establishing a DNA-binding site. These findings demonstrate that ChrN functions as a transcriptional repressor, modulating the cellular response of strain Z1 to Cr(VI) exposure.

Abstract Image

PadR 家族转录抑制因子调控肠杆菌 Z1 中铬酸盐外排转运体的转录
铬是一种普遍存在的有毒重金属,铬酸盐[Cr(VI)]具有高度诱变性和致癌性。在对六价铬具有抗性的菌株中,发现了六价铬外排蛋白 ChrA 的存在。然而,某些对六价铬具有抗性的菌株缺乏 ChrB(一种已知的调控因子)。在此,我们发现了一种 PadR 家族转录抑制因子 ChrN,它是肠杆菌 Z1(CCTCC NO: M 2019147)对六价铬反应的调节因子。ChrN 基因与 chrA 基因共转录,该操作子的转录表达受六价铬诱导。色氨酸荧光测定法和 Ni-NTA 纯化测定法都证明了 ChrN 蛋白与六价铬的结合能力。报告基因检测和电泳迁移检测验证了 ChrN 与 chrAN 操作子启动子之间的相互作用。保守组氨酸残基 His14 和 His50 的突变导致 ChrN 与 chrAN 操作子启动子失去结合。这一观察结果表明,这些残基对于建立 DNA 结合位点至关重要。这些研究结果表明,ChrN 可作为转录抑制因子,调节菌株 Z1 对六价铬暴露的细胞反应。
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来源期刊
Journal of Microbiology
Journal of Microbiology 生物-微生物学
CiteScore
5.70
自引率
3.30%
发文量
0
审稿时长
3 months
期刊介绍: Publishes papers that deal with research on microorganisms, including archaea, bacteria, yeasts, fungi, microalgae, protozoa, and simple eukaryotic microorganisms. Topics considered for publication include Microbial Systematics, Evolutionary Microbiology, Microbial Ecology, Environmental Microbiology, Microbial Genetics, Genomics, Molecular Biology, Microbial Physiology, Biochemistry, Microbial Pathogenesis, Host-Microbe Interaction, Systems Microbiology, Synthetic Microbiology, Bioinformatics and Virology. Manuscripts dealing with simple identification of microorganism(s), cloning of a known gene and its expression in a microbial host, and clinical statistics will not be considered for publication by JM.
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