title: Contribution of teg49 small RNA in the 5' upstream transcriptional region of sarA to virulence in Staphylococcus aureus.
abstract: High-throughput RNA sequencing technology has found the 5' untranslated region of sarA to contain two putative small RNAs (sRNAs), designated teg49 and teg48. Northern blot analysis disclosed that teg49 and teg48 were detectable within the P3-P1 and P1 sarA promoter regions, respectively. Focusing on teg49, we found that this sRNA, consisting of 196 nucleotides, is transcribed in the same direction as the sarA P3 transcript. The expression of both P3 and teg49 transcripts is dependent on sigB and cshA, which encodes a DEAD box RNA helicase. Within the sRNA teg49, there are two putative hairpin-loop structures, HP1 and HP2. Transversion mutation of the HP1 loop produced a smaller amount of sarA P3 and P2 transcripts and SarA protein than the corresponding HP1 stem and the HP2 stem and loop mutations, leading to lower RNAII transcription and derepression of aur transcription. The HP1 loop mutant also exhibited less biofilm formation than the parental and complemented strains. Complementation with shuttle plasmid pEPSA5 carrying teg49 was able to reestablish sarA P3 and P2 transcription and augment RNAII expression in the HP1 loop mutant. We thus conclude that teg49, embedded within the extended promoter regions of sarA, is modulated by sigB and cshA and plays an important trans-acting role in modulating the transcription and ensuing expression of sarA.
authors.affiliation: Department of Microbiology and Immunology, Geisel School of Medicine at Dartmouth, Hanover, New Hampshire, USA.
authors.forename: Samin
authors.initials: S
authors.lastname: Kim
authors.affiliation: Department of Microbiology and Immunology, Geisel School of Medicine at Dartmouth, Hanover, New Hampshire, USA.
authors.forename: Dindo
authors.initials: D
authors.lastname: Reyes
authors.affiliation: Service of Infectious Diseases, Geneva University Hospitals and Department of Microbiology and Molecular Medicine, University of Geneva, Geneva, Switzerland.
authors.forename: Marie
authors.initials: M
authors.lastname: Beaume
authors.affiliation: Genomic Research Lab, Services of Infectious Diseases, Geneva University Hospital, Geneva, Switzerland Patrice.francois@genomic.ch.
authors.forename: Patrice
authors.initials: P
authors.lastname: Francois
authors.affiliation: Department of Microbiology and Immunology, Geisel School of Medicine at Dartmouth, Hanover, New Hampshire, USA.
authors.forename: Ambrose
authors.initials: A
authors.lastname: Cheung
mesh_terms: D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D004252:DNA Mutational Analysis; D015964:Gene Expression Regulation, Bacterial; D005816:Genetic Complementation Test; D009690:Nucleic Acid Conformation; D017354:Point Mutation; D058727:RNA, Small Untranslated / Q000235:genetics / Q000378:metabolism*; D013211:Staphylococcus aureus / Q000235:genetics* / Q000472:pathogenicity*; D014157:Transcription Factors / Q000378:metabolism; D014158:Transcription, Genetic; D014774:Virulence
chemical_list: D001426:Bacterial Proteins; D058727:RNA, Small Untranslated; C492107:SarA protein, Staphylococcus aureus; D014157:Transcription Factors
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