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Journal of Bacteriology, December 2007, p. 8871-8879, Vol. 189, No. 24
0021-9193/07/$08.00+0     doi:10.1128/JB.01272-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Characterization of MazFSa, an Endoribonuclease from Staphylococcus aureus{triangledown}

Zhibiao Fu, Niles P. Donegan, Guido Memmi, and Ambrose L. Cheung*

Department of Microbiology and Immunology, Dartmouth Medical School, Hanover, New Hampshire 03755

Received 7 August 2007/ Accepted 2 October 2007

The mazEF homologs of Staphylococcus aureus, designated mazEFsa, have been shown to cotranscribe with the sigB operon under stress conditions. In this study, we showed that MazEFSa, as with their Escherichia coli counterparts, compose a toxin-antitoxin module wherein MazFSa leads to rapid cell growth arrest and loss in viable CFU upon overexpression. MazFSa is a novel sequence-specific endoribonuclease which cleaves mRNA to inhibit protein synthesis. Using ctpA mRNA as the model substrate both in vitro and in vivo, we demonstrated that MazFSa cleaves single-strand RNA preferentially at the 5' side of the first U or 3' side of the second U residue within the consensus sequences VUUV' (where V and V' are A, C, or G and may or may not be identical). Binding studies confirmed that the antitoxin MazESa binds MazFSa to form a complex to inhibit the endoribonuclease activity of MazFSa. Contrary to the system in E. coli, exposure to selected antibiotics augmented mazEFsa transcription, akin to what one would anticipate from the environmental stress response of the sigB system. These data indicate that the mazEF system of S. aureus differs from the gram-negative counterparts with respect to mRNA cleavage specificity and antibiotic stresses.

* Corresponding author. Mailing address: Department of Microbiology and Immunology, Dartmouth Medical School, Hanover, NH 03755. Phone: (603) 676-3350, ext. 2. Fax: (603) 676-3355. E-mail: Ambrose.Cheung{at}Dartmouth.edu

{triangledown} Published ahead of print on 12 October 2007.

Journal of Bacteriology, December 2007, p. 8871-8879, Vol. 189, No. 24
0021-9193/07/$08.00+0     doi:10.1128/JB.01272-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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