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Background  Research involving the cold shock gene  cspA  of the medically important bacterium  Staphylococcus aureus  is steadily increasing as the relationships between the activity of this gene at 37 °C and a spectrum of virulence factors (e.g., biofilm formation, capsule production) as well as stress-related genes (e.g., alkaline shock protein,  asp - 23  and the alternative sigma factor,  sigB ) are distinguished. Fundamental to each of these discoveries is defining the regulation of  cspA  and the production of its protein product CspA.    Results  In this paper, primer extension analysis was used to identify a transcriptional start point at 112 bp upstream of the initiation codon of the  cspA  coding sequence from  S. aureus  Newman RNA collected at 37 °C. Based on the location of the putative −10 and −35 sites as well as putative cold shock protein binding sites, a 192 bp sequence containing an 80 bp promoter + a 112 bp 5′ UTR was generated by polymerase chain reaction. The activity of this 192 bp sequence was confirmed in a pLL38 promoter:: xylE  reporter gene construct. In addition, Western blots were used to confirm the production of CspA at 37 °C and demonstrated that production of the protein was not constitutive but showed growth-dependent production with a significant increase at the 6 h time point.    Conclusions  The results presented identify another regulatory region for the cold shock gene  cspA  of  S. aureus  and show growth-dependent activity of both this  cspA  regulatory sequence, presented as a 192 bp sequence of promoter + 5′ UTR and the production of the CspA protein at 37 °C. The presence of two active transcription start points, a −112 bp sequence defined in this work and a second previously defined at −514 bp upstream of the  cspA  initiation codon, suggests the possibility of interactions between these two regions in the regulation of  cspA . The growth-dependent production of the cold shock protein CspA supports the availability of this protein to be a modulator of virulence and stress factor genes at 37 °C.

Growth-dependent activity of the cold shock cspA promoter + 5′ UTR and production of the protein CspA in Staphylococcus aureus Newman