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J Biotechnol. 2014 Nov 20;190:40-54. doi: 10.1016/j.jbiotec.2014.07.452. Epub 2014 Aug 6.

Transcriptional response of Corynebacterium glutamicum ATCC 13032 to hydrogen peroxide stress and characterization of the OxyR regulon.

Author information

1
Microbial Genomics and Biotechnology, CeBiTec, Bielefeld University, Bielefeld D-33615, Germany.
2
Microbial Genomics and Biotechnology, CeBiTec, Bielefeld University, Bielefeld D-33615, Germany; Technology Platform Genomics, CeBiTec, Bielefeld University, Bielefeld D-33615, Germany.
3
Microbial Genomics and Biotechnology, CeBiTec, Bielefeld University, Bielefeld D-33615, Germany; Technology Platform Genomics, CeBiTec, Bielefeld University, Bielefeld D-33615, Germany. Electronic address: joern.kalinowski@cebitec.uni-bielefeld.de.

Abstract

The aerobic soil bacterium Corynebacterium glutamicum ATCC 13032 has a remarkable natural resistance to hydrogen peroxide. A major player in hydrogen peroxide defense is the LysR type transcriptional regulator OxyR, homologs of which are present in a wide range of bacteria. In this study, the global transcriptional response of C. glutamicum to oxidative stress induced by hydrogen peroxide was examined using whole genome DNA microarrays, demonstrating the dynamic reaction of the regulatory networks. Deletion of oxyR resulted in an increased resistance of the C. glutamicum mutant to hydrogen peroxide. By performing DNA microarray hybridizations and RT-qPCR, differentially expressed genes were detected in the mutant. The direct control by OxyR was verified by electrophoretic mobility shift assays for 12 target regions. The results demonstrated that OxyR in C. glutamicum acts as a transcriptional repressor under non-stress conditions for a total of 23 genes. The regulated genes encode proteins related to oxidative stress response (e.g. katA), iron homeostasis (e.g. dps) and sulfur metabolism (e.g. suf cluster). Besides the regulator of the suf cluster, SufR, OxyR regulated the gene cg1695 encoding a putative transcriptional regulator, indicating the role of OxyR as a master regulator in defense against oxidative stress. Using a modified DNase footprint approach, the OxyR-binding sites in five target promoter regions, katA, cydA, hemH, dps and cg1292, were localized and in each upstream region at least two overlapping binding sites were found. The DNA regions protected by the OxyR protein are about 56bp in length and do not have evident sequence similarities. Still, by giving an insight in the H2O2 stimulon and extending the OxyR regulon this study considerably contributes to the understanding of the response of C. glutamicum to hydrogen peroxide-mediated oxidative stress.

KEYWORDS:

Corynebacterium glutamicum; DNase footprinting; Hydrogen peroxide; Next generation sequencing; Oxidative stress; OxyR; Transcriptional regulation

PMID:
25107507
DOI:
10.1016/j.jbiotec.2014.07.452
[Indexed for MEDLINE]

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