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The Gonococcal Transcriptome during Infection of the Lower Genital Tract in Women.

McClure R, Nudel K, Massari P, Tjaden B, Su X, Rice PA, Genco CA - PLoS ONE (2015)

Bottom Line: Gonorrhea is a highly prevalent disease resulting in significant morbidity worldwide, with an estimated 106 cases reported annually.A total of 140 genes were increased in expression during natural infection compared to growth in CDM, and 165 genes were decreased in expression.Large differences were found in gene expression profiles under each condition, particularly with genes involved in DNA and RNA processing, iron, transposase, pilin and lipoproteins.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Section of Infectious Diseases, Boston University School of Medicine, Boston, MA, United States of America; Department of Microbiology, Boston University School of Medicine, Boston, MA, United States of America.

ABSTRACT
Gonorrhea is a highly prevalent disease resulting in significant morbidity worldwide, with an estimated 106 cases reported annually. Neisseria gonorrhoeae, the causative agent of gonorrhea, colonizes and infects the human genital tract and often evades host immune mechanisms until successful antibiotic treatment is used. The alarming increase in antibiotic-resistant strains of N. gonorrhoeae, the often asymptomatic nature of this disease in women and the lack of a vaccine directed at crucial virulence determinants have prompted us to perform transcriptome analysis to understand gonococcal gene expression patterns during natural infection. We sequenced RNA extracted from cervico-vaginal lavage samples collected from women recently exposed to infected male partners and determined the complete N. gonorrhoeae transcriptome during infection of the lower genital tract in women. On average, 3.19% of total RNA isolated from female samples aligned to the N. gonorrhoeae NCCP11945 genome and 1750 gonococcal ORFs (65% of all protein-coding genes) were transcribed. High expression in vivo was observed in genes encoding antimicrobial efflux pumps, iron response, phage production, pilin structure, outer membrane structures and hypothetical proteins. A parallel analysis was performed using the same strains grown in vitro in a chemically defined media (CDM). A total of 140 genes were increased in expression during natural infection compared to growth in CDM, and 165 genes were decreased in expression. Large differences were found in gene expression profiles under each condition, particularly with genes involved in DNA and RNA processing, iron, transposase, pilin and lipoproteins. We specifically interrogated genes encoding DNA binding regulators and iron-scavenging proteins, and identified increased expression of several iron-regulated genes, including tbpAB and fbpAB, during infection in women as compared to growth in vitro, suggesting that during infection of the genital tract in women, the gonococcus is exposed to an iron deplete environment. Collectively, we demonstrate that a large portion of the gonococcal genome is expressed and regulated during mucosal infection including genes involved in regulatory functions and iron scavenging.

No MeSH data available.


Related in: MedlinePlus

Subject-specific comparisons between transcriptomes of N. gonorrhoeae in natural infection vs. growth in CDM.N. gonorrhoeae transcriptomes observed for each subject compared with transcriptomes of corresponding infecting strains grown in CDM. Genes that showed at least a 3-fold change in expression were categorized based on gene function. (A) Functional enrichment of genes showing at least a 3-fold change in expression during infection of Subject 1 compared to growth of the infecting strain in CDM. Gene categories are shown in the y-axis and ratio of enrichment is shown on the x-axis. Blue bars indicate functional enrichment of genes showing increased expression during natural infection in vivo compared to growth in CDM. Red bars indicate functional enrichment of genes showing decreased expression during natural infection in vivo compared to growth in CDM. (B) Identical analysis for Subject 2 (C) Identical analysis for Subject 3.
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pone.0133982.g003: Subject-specific comparisons between transcriptomes of N. gonorrhoeae in natural infection vs. growth in CDM.N. gonorrhoeae transcriptomes observed for each subject compared with transcriptomes of corresponding infecting strains grown in CDM. Genes that showed at least a 3-fold change in expression were categorized based on gene function. (A) Functional enrichment of genes showing at least a 3-fold change in expression during infection of Subject 1 compared to growth of the infecting strain in CDM. Gene categories are shown in the y-axis and ratio of enrichment is shown on the x-axis. Blue bars indicate functional enrichment of genes showing increased expression during natural infection in vivo compared to growth in CDM. Red bars indicate functional enrichment of genes showing decreased expression during natural infection in vivo compared to growth in CDM. (B) Identical analysis for Subject 2 (C) Identical analysis for Subject 3.

Mentions: Regulated genes were those that showed a difference in expression of at least 3-fold (up or down) in natural infection compared to growth in CDM. On average, 397 gonococcal genes, representing 20% of the genome, were regulated in each subject. Functional enrichment analysis, performed on genes showing differences in expression during natural infection compared to growth in CDM, showed enrichment of genes encoding for tRNA, iron, membrane, transposase and lipoproteins among genes whose expression was increased in vivo compared to growth in CDM. In contrast, metabolism, growth and housekeeping functions were enriched in genes showing decreased expression in vivo compared to CDM (Fig 3). Subject-specific gene expression patterns appeared overall remarkably similar, although subtle differences were also noted between subjects. For example, in the comparison between in vivo gene expression and growth in CDM, pilin-related genes were enriched among genes showing increased expression in Subject 2, enriched among genes showing decreased expression in Subject 3 and approximately equal Subject 1. A similar variability was observed for stress response genes, where enrichment among genes with increased expression was observed in Subject 3 but not in Subject 1 or 2. Finally, genes encoding lipoproteins were enriched among genes with increased expression in Subject 1 but not in Subjects 2 or 3. Overall, we observed general trends and a core set of genes differentially expressed in all subjects regardless of the specifics of infection. However, the subtle differences in regulation and expression of gonococcal genes, for example those involved in pilin and stress response, indicate that the genital tract of each subject was unique.


The Gonococcal Transcriptome during Infection of the Lower Genital Tract in Women.

McClure R, Nudel K, Massari P, Tjaden B, Su X, Rice PA, Genco CA - PLoS ONE (2015)

Subject-specific comparisons between transcriptomes of N. gonorrhoeae in natural infection vs. growth in CDM.N. gonorrhoeae transcriptomes observed for each subject compared with transcriptomes of corresponding infecting strains grown in CDM. Genes that showed at least a 3-fold change in expression were categorized based on gene function. (A) Functional enrichment of genes showing at least a 3-fold change in expression during infection of Subject 1 compared to growth of the infecting strain in CDM. Gene categories are shown in the y-axis and ratio of enrichment is shown on the x-axis. Blue bars indicate functional enrichment of genes showing increased expression during natural infection in vivo compared to growth in CDM. Red bars indicate functional enrichment of genes showing decreased expression during natural infection in vivo compared to growth in CDM. (B) Identical analysis for Subject 2 (C) Identical analysis for Subject 3.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4526530&req=5

pone.0133982.g003: Subject-specific comparisons between transcriptomes of N. gonorrhoeae in natural infection vs. growth in CDM.N. gonorrhoeae transcriptomes observed for each subject compared with transcriptomes of corresponding infecting strains grown in CDM. Genes that showed at least a 3-fold change in expression were categorized based on gene function. (A) Functional enrichment of genes showing at least a 3-fold change in expression during infection of Subject 1 compared to growth of the infecting strain in CDM. Gene categories are shown in the y-axis and ratio of enrichment is shown on the x-axis. Blue bars indicate functional enrichment of genes showing increased expression during natural infection in vivo compared to growth in CDM. Red bars indicate functional enrichment of genes showing decreased expression during natural infection in vivo compared to growth in CDM. (B) Identical analysis for Subject 2 (C) Identical analysis for Subject 3.
Mentions: Regulated genes were those that showed a difference in expression of at least 3-fold (up or down) in natural infection compared to growth in CDM. On average, 397 gonococcal genes, representing 20% of the genome, were regulated in each subject. Functional enrichment analysis, performed on genes showing differences in expression during natural infection compared to growth in CDM, showed enrichment of genes encoding for tRNA, iron, membrane, transposase and lipoproteins among genes whose expression was increased in vivo compared to growth in CDM. In contrast, metabolism, growth and housekeeping functions were enriched in genes showing decreased expression in vivo compared to CDM (Fig 3). Subject-specific gene expression patterns appeared overall remarkably similar, although subtle differences were also noted between subjects. For example, in the comparison between in vivo gene expression and growth in CDM, pilin-related genes were enriched among genes showing increased expression in Subject 2, enriched among genes showing decreased expression in Subject 3 and approximately equal Subject 1. A similar variability was observed for stress response genes, where enrichment among genes with increased expression was observed in Subject 3 but not in Subject 1 or 2. Finally, genes encoding lipoproteins were enriched among genes with increased expression in Subject 1 but not in Subjects 2 or 3. Overall, we observed general trends and a core set of genes differentially expressed in all subjects regardless of the specifics of infection. However, the subtle differences in regulation and expression of gonococcal genes, for example those involved in pilin and stress response, indicate that the genital tract of each subject was unique.

Bottom Line: Gonorrhea is a highly prevalent disease resulting in significant morbidity worldwide, with an estimated 106 cases reported annually.A total of 140 genes were increased in expression during natural infection compared to growth in CDM, and 165 genes were decreased in expression.Large differences were found in gene expression profiles under each condition, particularly with genes involved in DNA and RNA processing, iron, transposase, pilin and lipoproteins.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Section of Infectious Diseases, Boston University School of Medicine, Boston, MA, United States of America; Department of Microbiology, Boston University School of Medicine, Boston, MA, United States of America.

ABSTRACT
Gonorrhea is a highly prevalent disease resulting in significant morbidity worldwide, with an estimated 106 cases reported annually. Neisseria gonorrhoeae, the causative agent of gonorrhea, colonizes and infects the human genital tract and often evades host immune mechanisms until successful antibiotic treatment is used. The alarming increase in antibiotic-resistant strains of N. gonorrhoeae, the often asymptomatic nature of this disease in women and the lack of a vaccine directed at crucial virulence determinants have prompted us to perform transcriptome analysis to understand gonococcal gene expression patterns during natural infection. We sequenced RNA extracted from cervico-vaginal lavage samples collected from women recently exposed to infected male partners and determined the complete N. gonorrhoeae transcriptome during infection of the lower genital tract in women. On average, 3.19% of total RNA isolated from female samples aligned to the N. gonorrhoeae NCCP11945 genome and 1750 gonococcal ORFs (65% of all protein-coding genes) were transcribed. High expression in vivo was observed in genes encoding antimicrobial efflux pumps, iron response, phage production, pilin structure, outer membrane structures and hypothetical proteins. A parallel analysis was performed using the same strains grown in vitro in a chemically defined media (CDM). A total of 140 genes were increased in expression during natural infection compared to growth in CDM, and 165 genes were decreased in expression. Large differences were found in gene expression profiles under each condition, particularly with genes involved in DNA and RNA processing, iron, transposase, pilin and lipoproteins. We specifically interrogated genes encoding DNA binding regulators and iron-scavenging proteins, and identified increased expression of several iron-regulated genes, including tbpAB and fbpAB, during infection in women as compared to growth in vitro, suggesting that during infection of the genital tract in women, the gonococcus is exposed to an iron deplete environment. Collectively, we demonstrate that a large portion of the gonococcal genome is expressed and regulated during mucosal infection including genes involved in regulatory functions and iron scavenging.

No MeSH data available.


Related in: MedlinePlus