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  1. Article ; Online: The novel protein ScrA acts through the SaeRS two-component system to regulate virulence gene expression in Staphylococcus aureus.

    Wittekind, Marcus A / Frey, Andrew / Bonsall, Abigail E / Briaud, Paul / Keogh, Rebecca A / Wiemels, Richard E / Shaw, Lindsey N / Carroll, Ronan K

    Molecular microbiology

    2022  Volume 117, Issue 5, Page(s) 1196–1212

    Abstract: Staphylococcus aureus is a Gram-positive commensal that can also cause a variety of infections in humans. S. aureus virulence factor gene expression is under tight control by a complex regulatory network, which includes, sigma factors, sRNAs, and two- ... ...

    Abstract Staphylococcus aureus is a Gram-positive commensal that can also cause a variety of infections in humans. S. aureus virulence factor gene expression is under tight control by a complex regulatory network, which includes, sigma factors, sRNAs, and two-component systems (TCS). Previous work in our laboratory demonstrated that overexpression of the sRNA tsr37 leads to an increase in bacterial aggregation. Here, we demonstrate that the clumping phenotype is dependent on a previously unannotated 88 amino acid protein encoded within the tsr37 sRNA transcript (which we named ScrA for S. aureus clumping regulator A). To investigate the mechanism of action of ScrA we performed proteomics and transcriptomics in a ScrA overexpressing strain and show that a number of surface adhesins are upregulated, while secreted proteases are downregulated. Results also showed upregulation of the SaeRS TCS, suggesting that ScrA is influencing SaeRS activity. Overexpression of ScrA in a saeR mutant abrogates the clumping phenotype confirming that ScrA functions via the Sae system. Finally, we identified the ArlRS TCS as a positive regulator of scrA expression. Collectively, our results show that ScrA is an activator of the SaeRS system and suggests that ScrA may act as an intermediary between the ArlRS and SaeRS systems.
    MeSH term(s) Bacterial Proteins/genetics ; Bacterial Proteins/metabolism ; Gene Expression ; Gene Expression Regulation, Bacterial/genetics ; Humans ; Protein Kinases/metabolism ; RNA, Small Untranslated/metabolism ; Staphylococcal Infections ; Staphylococcus aureus/metabolism ; Transcription Factors/genetics ; Transcription Factors/metabolism ; Virulence/genetics
    Chemical Substances Bacterial Proteins ; RNA, Small Untranslated ; Transcription Factors ; Protein Kinases (EC 2.7.-)
    Language English
    Publishing date 2022-04-15
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, N.I.H., Extramural
    ZDB-ID 619315-8
    ISSN 1365-2958 ; 0950-382X
    ISSN (online) 1365-2958
    ISSN 0950-382X
    DOI 10.1111/mmi.14901
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: Global Annotation, Expression Analysis, and Stability of Candidate sRNAs in Group B Streptococcus.

    Keogh, Rebecca A / Spencer, Brady L / Sorensen, Hailee M / Zapf, Rachel L / Briaud, Paul / Bonsall, Abigail E / Doran, Kelly S / Carroll, Ronan K

    mBio

    2021  Volume 12, Issue 6, Page(s) e0280321

    Abstract: Small, noncoding RNAs (sRNAs) are being increasingly identified as important regulatory molecules in prokaryotes. Due to the prevalence of next-generation sequencing-based techniques, such as RNA sequencing (RNA-seq), there is potential for increased ... ...

    Abstract Small, noncoding RNAs (sRNAs) are being increasingly identified as important regulatory molecules in prokaryotes. Due to the prevalence of next-generation sequencing-based techniques, such as RNA sequencing (RNA-seq), there is potential for increased discovery of sRNAs within bacterial genomes; however, these elements are rarely included in annotation files. Consequently, expression values for sRNAs are omitted from most transcriptomic analyses, and mechanistic studies have lagged behind those of protein regulators in numerous bacteria. Two previous studies have identified sRNAs in the human pathogen group B Streptococcus (GBS). Here, we utilize the data from these studies to create updated genome annotation files for the model GBS strains NEM316 and COH1. Using the updated COH1 annotation file, we reanalyze publicly available GBS RNA-seq whole-transcriptome data from GenBank to monitor GBS sRNA expression under a variety of conditions and genetic backgrounds. This analysis generated expression values for 232 putative sRNAs that were overlooked in previous transcriptomic analyses in 21 unique comparisons. To demonstrate the utility of these data, we identify an sRNA that is upregulated during vaginal colonization and demonstrate that overexpression of this sRNA leads to increased bacterial invasion into host epithelial cells. Finally, to monitor RNA degradation, we perform a transcript stability assay to identify highly stable sRNAs and compare stability profiles of sRNA- and protein-coding genes. Collectively, these data provide a wealth of transcriptomic data for putative sRNAs in GBS and a platform for future mechanistic studies.
    MeSH term(s) Gene Expression Profiling ; Gene Expression Regulation, Bacterial ; Genome, Bacterial ; Humans ; RNA Stability ; RNA, Bacterial/chemistry ; RNA, Bacterial/genetics ; RNA, Bacterial/metabolism ; RNA, Small Untranslated/chemistry ; RNA, Small Untranslated/genetics ; RNA, Small Untranslated/metabolism ; Streptococcal Infections/microbiology ; Streptococcus agalactiae/chemistry ; Streptococcus agalactiae/genetics ; Streptococcus agalactiae/metabolism
    Chemical Substances RNA, Bacterial ; RNA, Small Untranslated
    Language English
    Publishing date 2021-11-02
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural
    ZDB-ID 2557172-2
    ISSN 2150-7511 ; 2161-2129
    ISSN (online) 2150-7511
    ISSN 2161-2129
    DOI 10.1128/mBio.02803-21
    Database MEDical Literature Analysis and Retrieval System OnLINE

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