Abstract
The pathogenesis of Staphylococcus aureus, from local infections to systemic dissemination, is mediated by a battery of virulence factors that are regulated by intricate mechanisms, which include regulatory proteins and small RNAs (sRNAs) as key regulatory molecules. We have investigated the involvement of sRNA RsaF, in the regulation of pathogenicity genes hyaluronate lyase (hysA) and serine proteaselike protein D (splD), by employing S. aureus strains with disruption and overexpression of rsaF. Staphylococcus aureus strain with disruption of rsaF exhibited marked down-regulation of hysA transcripts by 0.2 to 0.0002 fold, and hyaluronate lyase activity by 0.2–0.1 fold, as well as increased biofilm formation, during growth from log phase to stationery phase. These mutants also displayed down-regulation of splD transcripts by 0.8 to 0.005 fold, and reduced activity of multiple proteases by zymography. Conversely, overexpression of rsaF resulted in a 2- to 4- fold increase in hysA mRNA levels and hyaluronidase activity. Both hysA and splD mRNAs demonstrated an increased stability in RsaF+ strains. In silico RNA-RNA interaction indicated a direct base pairing of RsaF with hysA and splD mRNAs, which was established in electrophoretic mobility shift assays. The findings demonstrate a positive regulatory role for small RNA RsaF in the expression of the virulence factors, HysA and SplD.
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Acknowledgements
NP was supported by BSR Fellowship in Science F.4-1/2006-(BSR)/7-128/2007(BSR) by University Grant Commission, Government of India, New Delhi.
The authors thank Professors Timothy Foster and Ian. R. Monk, Trinity College, Dublin for S. aureus and E. coli strains. We thank Professor Cristiane Wolz, University of Tubingen, Tubingen, Germany for supplying pMAD plasmid. The pCN40 plasmid was obtained from Network on antimicrobial resistance in Staphylococcus aureus (NARSA) program supported under NIAD, NIH contract No. HHSN272200700055C. We thank DBT-MSUB-ILSPARE, Dr. Vikram Sarabhai Science Block, Maharaja Sayajirao University of Baroda for the Confocal microscopy facility.
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Patel, N., Nair, M. The small RNA RsaF regulates the expression of secreted virulence factors in Staphylococcus aureus Newman. J Microbiol. 59, 920–930 (2021). https://doi.org/10.1007/s12275-021-1205-6
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DOI: https://doi.org/10.1007/s12275-021-1205-6