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Transcriptional effects of overexpression of the mtrCDE operon in Neisseria gonorrhoeae due to a cis-acting point mutation

ABSTRACT: A whole transcriptome comparison of N. gonorrhoeae strain FA19, which bears a wild-type mtr locus, and FA19mtr120, which contains a C-to-T transition mutation 120 base pairs upstream of the mtrC start codon that generates a second promoter for mtrCDE transcription, was performed to determine the effects of high-level expression of the mtrCDE efflux pump operon on global transcription within the gonococcus.

ORGANISM(S): Neisseria gonorrhoeae FA19

PROVIDER: GSE47048 | GEO | 2013/05/18

SECONDARY ACCESSION(S): PRJNA203368

REPOSITORIES: GEO

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Transcriptional effects of overexpression of the mtrCDE operon in Neisseria gonorrhoeae due to a cis-acting point mutation

Project description:A whole transcriptome comparison of N. gonorrhoeae strain FA19, which bears a wild-type mtr locus, and FA19mtr120, which contains a C-to-T transition mutation 120 base pairs upstream of the mtrC start codon that generates a second promoter for mtrCDE transcription, was performed to determine the effects of high-level expression of the mtrCDE efflux pump operon on global transcription within the gonococcus. Examination of the transcriptomes of N. gonorrhoeae wild-type strain FA19 and the isogenic FA19mtr120 mutant to determine the effect of overexpression of mtrCDE on gonococcal cells grown under laboratory conditions. Illumina HiSeq-2000 next generation sequencing was used to sequence the transcriptomes of each strain. Reads were mapped against the N. gonorrhoeae FA1090 genome (NCBI accession number NC_002946) because at the time this experiment was run the FA19 genome was incomplete.

2013-05-18 | E-GEOD-47048 | biostudies-arrayexpress

MpeR regulation in Neisseria gonorrhoeae (F19) through an iron-responsive mechanism

Project description:Previous studies have shown that the MpeR transcriptional regulator produced by Neisseria gonorrhoeae represses expression of mtrF, which encodes a putative inner membrane protein that works with the MtrC-MtrD-MtrE efflux pump to allow gonococci to resist high levels of multiple hydrophobic antimicrobials. Regulation of mpeR has been reported to occur by an iron-dependent mechanism involving Fur (Ferric uptake regulator). Collectively, these observations suggest the presence of an interconnected regulatory system in gonococci that modulates expression of drug efflux pump protein-encoding genes in an iron-responsive manner. Herein, we describe this connection and report that levels of gonococcal resistance to a substrate of the mtrCDE-encoded efflux pump can be modulated by MpeR and the availability of free iron. Using microarray analysis, we found that the mtrR gene, which encodes the direct transcriptional repressor (MtrR) of mtrCDE, is an MpeR-repressed determinant in the late-logarithmic phase of growth when free iron levels would be reduced due to bacterial consumption. MpeR-mediated repression of mtrR appeared to be direct, as judged by DNA-binding analyses, and was enhanced by conditions of iron-limitation, which resulted in increased expression of the mtrCDE efflux pump operon. Taken together, our results indicate that both genetic and physiologic parameters can influence expression of the mtr efflux system and that these can modulate levels of gonococcal susceptibility to efflux pump substrates.

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MpeR regulation in Neisseria gonorrhoeae (F19) through an iron-responsive mechanism

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