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Enhancement of the pathogenicity of Staphylococcus aureus strain Newman by a small noncoding RNA SprX1

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Abstract

The pathogenesis of Staphylococcus aureus from local infection to systemic dissemination involves a range of virulence factors including structural and secreted products. Among various control mechanisms, small noncoding RNAs are involved in the regulation of multiple pathogenicity factors in S. aureus. The sRNA SprX which is encoded in the pathogenicity island of methicillin-susceptible S. aureus strain Newman and was shown to influence antibiotic resistance previously, upregulated the expression of virulence genes, especially the cell wall-associated clumping factor B (ClfB) and delta hemolysin (Hld). Bioinformatic analysis revealed several multiple mRNAs associated with pathogenicity as targets for SprX1, one of the three copies of sprX. Both overexpression and chromosomal disruption of sprX1 supported the scheme of upregulation of clfB and hld expression. Altered expression of SprX1 altered the levels of Hld and ClfB mRNAs, hemolysis, clumping of cells, biofilm formation by plate adhesion studies and confocal microscopic analysis as well as infection pathology of modified strains in mice models. ClfB and Hld mRNAs interacted directly with SprX1 in in vitro assays. Increased level of the regulatory RNA, namely RNAIII, that comprises Hld mRNA and also regulates the biofilm formation, indicates that SprX1 may also function through RNAIII for regulating virulence factors. An immunodominant protein, antigen A, was downregulated by SprX1 in two-dimensional electrophoresis. Taken together, these results signify the role of sRNA SprX in the pathogenicity of S. aureus Newman.

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Acknowledgments

We thank Professors Timothy Foster and Ian. R. Monk, Trinity College, Dublin, Abraham. L. Sonenshein, Tufts University, Boston, for providing shuttle plasmids, S. aureus and E. coli strains. The pCN40 vector was obtained through the Network on Antimicrobial Resistance in Staphylococcus aureus (NARSA) program supported under NIAD, NIH Contract No. HHSN272200700055C. MK was supported by a research fellowship (F.4-1/2006(BSR)/7-128/2007) by University Grant Commission, Government of India. The initial phase of this work was partially funded by a grant from the Department of Biotechnology, India (BT/PR/P0056/AGR/36/29/2007).

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  1. Department of Microbiology and Biotechnology Centre, The Maharaja Sayajirao University of Baroda, Vadodara, Gujarat, 390002, India

    Manikandan Kathirvel, Hasmatbanu Buchad & Mrinalini Nair

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  1. Manikandan Kathirvel
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The study protocols for the experimental use of mice were carried out in strict accordance with and approved by Committee for the Purpose of Control and Supervision on Experiments on Animals (CPCSEA), Government of India (Approval number 938/PO/a/06/CPCSEA).

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Kathirvel, M., Buchad, H. & Nair, M. Enhancement of the pathogenicity of Staphylococcus aureus strain Newman by a small noncoding RNA SprX1. Med Microbiol Immunol 205, 563–574 (2016). https://doi.org/10.1007/s00430-016-0467-9

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