Identification of key determinants of<i>Staphylococcus aureus</i>vaginal colonization

Deng, Liwen; Schilcher, Katrin; Burcham, Lindsey R.; Kwieciński, Jakub; Johnson, Paige; Head, Steven R.; Heinrichs, David E.; Horswill, Alexander R.; Doran, Kelly S.

doi:10.1101/761841

Cited by 3 publications

(4 citation statements)

References 101 publications

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“…We did not observe background green fluorescence in naïve mice (Fig. S1B, C, D), which coincides with previous experiments performed with GBS and S. aureus (26,27). The presence of fluorescent E. faecalis in the cervix and uterus shows that E. faecalis can move from the vaginal lumen to the superior organs of the female reproductive tract.…”

Section: E Faecalis Colonization Of the Female Reproductive Tractsupporting

confidence: 90%

“…1B, C). This level and range of recovered E. faecalis CFU is similar to what we have observed using this mouse model for GBS and S. aureus vaginal colonization (26,27). To visualize E. faecalis within the murine reproductive tract, mice were inoculated with either WT E. faecalis V583 or a V583 strain expressing green fluorescent protein (GFP) (28).…”

Section: E Faecalis Colonization Of the Female Reproductive Tractsupporting

confidence: 81%

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Genome-Wide Mutagenesis Identifies Factors Involved in Enterococcus faecalis Vaginal Adherence and Persistence

et al. 2020

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Enterococcus faecalis is a Gram-positive commensal bacterium native to the gastrointestinal tract and an opportunistic pathogen of increasing clinical concern. E. faecalis also colonizes the female reproductive tract and reports suggest vaginal colonization increases following antibiotic treatment or in patients with aerobic vaginitis. Currently, little is known about specific factors that promote E. faecalis vaginal colonization and subsequent infection. We modified an established mouse vaginal colonization model to explore E. faecalis vaginal carriage and demonstrate that both vancomycin resistant and sensitive strains colonize the murine vaginal tract. Following vaginal colonization, we observed E. faecalis in vaginal, cervical and uterine tissue. A mutant lacking endocarditis- and biofilm-associated pili (Ebp) exhibited a decreased ability to associate with human vaginal and cervical cells in vitro, but did not contribute to colonization in vivo. Thus, we screened a low-complexity transposon (Tn) mutant library to identify novel genes important for E. faecalis colonization and persistence in the vaginal tract. This screen revealed 383 mutants that were underrepresented during vaginal colonization at 1, 5 and 8 days post-inoculation compared to growth in culture medium. We confirmed that mutants deficient in ethanolamine catabolism or in the type VII secretion system were attenuated in persisting during vaginal colonization. These results reveal the complex nature of vaginal colonization and suggest that multiple factors contribute to E. faecalis persistence in the reproductive tract. IMPORTANCE Despite increasing prevalence and association of E. faecalis with aerobic vaginitis, essentially nothing is known about the bacterial factors that influence E. faecalis vaginal colonization. We have adapted an animal model of vaginal colonization that supports colonization of multiple E. faecalis strains. Additionally, we determined that ethanolamine utilization and type VII secretion system genes contribute to vaginal colonization and persistence. Identification of factors important for vaginal colonization and persistence provides potential targets for the development of therapeutics. This study is the first to identify key determinants that promote vaginal colonization by E. faecalis, which may represent an important reservoir for antibiotic resistant enterococci.

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Section: E Faecalis Colonization Of the Female Reproductive Tractsupporting

confidence: 90%

Section: E Faecalis Colonization Of the Female Reproductive Tractsupporting

confidence: 81%

Genome-Wide Mutagenesis Identifies Factors Involved in Enterococcus faecalis Vaginal Adherence and Persistence

et al. 2020

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“…Recently, two studies have utilized RNA-Seq to examine the transcriptional response of S. aureus in vivo . Work by Ibberson and Whiteley examined the S. aureus transcriptome in the lung of patients with cystic fibrosis (CF) ( 14 ), while Deng et al examined the S. aureus transcriptome during vaginal colonization ( 15 ). The RNA-Seq data sets for these two studies were not yet available at the time that we performed the original search of the GEO database (for our global sRNA expression analysis described above).…”

Section: Resultsmentioning

confidence: 99%

“…In vivo RNA-Seq data sets were acquired from GEO under accession number SRP222773 for CF data and accession number SRP229518 for vaginal data ( 14 , 15 ). For the CF lung data, sRNA expression values (in RPKM) from nine clinical isolate samples were averaged and compared to the averaged sRNA expression values from nine S. aureus cultures grown in chemically defined media (CDM) as well as to those from nine samples from synthetic cystic fibrosis media (SCFM).…”

Section: Methodsmentioning

confidence: 99%

Reading between the Lines: Utilizing RNA-Seq Data for Global Analysis of sRNAs in Staphylococcus aureus

Sorensen

Keogh

Wittekind

et al. 2020

mSphere

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Regulatory small RNAs (sRNAs) are known to play important roles in the Gram-positive bacterial pathogen Staphylococcus aureus; however, their existence is often overlooked, primarily because sRNA genes are absent from genome annotation files. Consequently, transcriptome sequencing (RNA-Seq)-based experimental approaches, performed using standard genome annotation files as a reference, have likely overlooked data for sRNAs. Previously, we created an updated S. aureus genome annotation file, which included annotations for 303 known sRNAs in USA300. Here, we utilized this updated reference file to reexamine publicly available RNA-Seq data sets in an attempt to recover lost information on sRNA expression, stability, and potential to encode peptides. First, we used transcriptomic data from 22 studies to identify how the expression of 303 sRNAs changed under 64 different experimental conditions. Next, we used RNA-Seq data from an RNA stability assay to identify highly stable/unstable sRNAs. We went on to reanalyze a ribosome profiling (Ribo-seq) data set to identify sRNAs that have the potential to encode peptides and to experimentally confirm the presence of three of these peptides in the USA300 background. Interestingly, one of these sRNAs/peptides, encoded at the tsr37 locus, influences the ability of S. aureus cells to autoaggregate. Finally, we reexamined two recently published in vivo RNA-Seq data sets, from the cystic fibrosis (CF) lung and a murine vaginal colonization study, and identified 29 sRNAs that may play a role in vivo. Collectively, these results can help inform future studies of these important regulatory elements in S. aureus and highlight the need for ongoing curating and updating of genome annotation files. IMPORTANCE Regulatory small RNAs (sRNAs) are a class of RNA molecules that are produced in bacterial cells but that typically do not encode proteins. Instead, they perform a variety of critical functions within the cell as RNA. Most bacterial genomes do not include annotations for sRNA genes, and any type of analysis that is performed using a bacterial genome as a reference will therefore overlook data for sRNAs. In this study, we reexamined hundreds of previously generated S. aureus RNA-Seq data sets and reanalyzed them to generate data for sRNAs. To do so, we utilized an updated S. aureus genome annotation file, previously generated by our group, which contains annotations for 303 sRNAs. The data generated (which were previously discarded) shed new light on sRNAs in S. aureus, most of which are unstudied, and highlight certain sRNAs that are likely to play important roles in the cell.

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Group B Streptococcus Adaptation Promotes Survival in a Hyperinflammatory Diabetic Wound Environment

Keogh

Haeberle

Langouët-Astrié

et al. 2022

Preprint

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Diabetic wounds have poor healing outcomes due to the presence of numerous pathogens and a dysregulated immune response. Group B Streptococcus (GBS) is commonly isolated from diabetic wound infections, but the mechanisms of GBS virulence during these infections have not been investigated. Here, we develop a murine model of GBS diabetic wound infection, and using dual RNA-sequencing, demonstrate that GBS infection triggers an inflammatory response. GBS adapts to this hyperinflammatory environment by upregulating virulence factors including those known to be regulated by the the two-component system covRS, such as the surface protein pbsP, and the cyl operon which is responsible for hemolysin/pigmentation production. We recover hyper-pigmented/hemolytic GBS colonies from the murine diabetic wound which we determined encode mutations in covR. We further demonstrate that GBS mutants in cylE and pbsP are attenuated in the diabetic wound. This foundational study provides insight into the pathogenesis of GBS diabetic wound infections.

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Identification of key determinants ofStaphylococcus aureusvaginal colonization

Cited by 3 publications

References 101 publications

Genome-Wide Mutagenesis Identifies Factors Involved in Enterococcus faecalis Vaginal Adherence and Persistence

Genome-Wide Mutagenesis Identifies Factors Involved in Enterococcus faecalis Vaginal Adherence and Persistence

Reading between the Lines: Utilizing RNA-Seq Data for Global Analysis of sRNAs in Staphylococcus aureus

Group B Streptococcus Adaptation Promotes Survival in a Hyperinflammatory Diabetic Wound Environment

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