Unknown,Transcriptomics,Genomics,Proteomics

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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. 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.

ORGANISM(S): Neisseria gonorrhoeae FA19

SUBMITTER: Gregory Doho

PROVIDER: E-GEOD-47048 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:The emergence of fully antimicrobial resistant Neisseria gonorrhoeae has led global public health agencies to identify a critical need for next generation anti-gonococcal pharmaceuticals. The development and success of these compounds will rely upon valid pre-clinical models of gonorrhoeae infection. We recently developed and reported the first model of upper genital tract gonococcal infection. During initial characterization, we observed significant reproductive cycle-based variation in infection outcome. When uterine infection occurred in the diestrus phase, there was significantly greater pathology than during estrus phase. The aim of this study was to evaluate transcriptional profiles of infected uterine tissue from mice in either estrus or diestrus phase in order to elucidate possible mechanisms for these differences. Genes and biological pathways with phase-independent induction during infection showed a chemokine dominant cytokine response to Neisseria gonorrhoeae. Despite general induction being phase-independent, this common anti-gonococcal response demonstrated greater induction during diestrus phase infection. Greater activity of granulocyte adhesion and diapedesis regulators during diestrus infection, particularly in chemokines and diapedesis regulators, was also shown. In addition to a greater induction of the common anti-gonococcal response, Gene Set Enrichment Analysis (GSEA) identified a diestrus-specific induction of type-1 interferon signaling pathways. This transcriptional analysis of murine uterine gonococcal infection during distinct points in the natural reproductive cycle provided evidence for a common anti-gonococcal response characterized by significant induction of granulocyte chemokine expression and high proinflammatory mediators. The basic biology of this host response to N. gonorrhoeae in estrus and diestrus is similar at the pathway level, but varies drastically in magnitude. Overlaying this, we observed type-1 interferon induction specifically in diestrus infection where greater pathology is observed. This supports recent work suggesting this pathway has a significant, possibly host-detrimental, function in gonococcal infection. Together these findings lay the groundwork for further examination of the role of interferons in gonococcal infection. Additionally, this work enables the implementation of the diestrus uterine infection model using the newly characterized host response as a marker of pathology and its prevention as a correlate of candidate vaccine efficacy and ability to protect against the devastating consequences of N. gonorrhoeae-associated sequelae. Murine microarrays were used to examine transcriptional differences underlying significantly different phenotypes associated with uterine N. gonorrhoeae infection in the estrus versus diestrus phases of the natural reproductive cycle.

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

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