1887

Abstract

has several extracellular proteases with proposed roles in virulence. SspA (serine protease), SspB (cysteine protease) and Aur (metalloprotease) have been characterized previously and SspA and SspB were found to be cotranscribed. The coding region for the cysteine protease ScpA has been identified and characterized. It is in a probable bi-cistronic operon with located immediately upstream of a coding region for a 108 aa protein that is a specific inhibitor of ScpA. Using primer extension analysis promoters have been mapped and it was found that is the only sigma factor involved in the transcription of , and . The transcription of all the genes occurs maximally at post-exponential phase, being positively regulated by (accessory gene regulator) and negatively regulated by (staphylococcal accessory regulator). Furthermore represses transcription from the and operons similarly to the previously shown effect on [ Horsburgh, M., Aish, J., White, I., Shaw, L., Lithgow, J. & Foster, S. (2002). , 5457–5467 ]. Using mutations in each protease gene the proteolytic cascade of activation has been analysed. Aur, SspA, SspB and ScpA are all produced as zymogens, activated by proteolytic cleavage. Although the metalloprotease, Aur, does catalyse activation of the SspA zymogen, it is not the sole agent capable of conducting this process. Site-directed mutagenesis revealed that Aur is not capable of undergoing auto-proteolysis to achieve activation. The cysteine protease, ScpA, appears to reside outside this cascade of activation, as mature ScpA was observed in the , and mutant strains. Using a mouse abscess model, it has been shown that insertional inactivation of or results in significant attenuation of virulence, whilst mutations in or do not. It is likely the attenuation observed in the strain is due to polarity on the gene.

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2004-01-01
2019-10-23
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