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Background  The highly conserved macromolecular synthesis operon (MMSO) contains both  dnaG  (primase) and  sigA  (primary sigma factor). However, in previously evaluated gram-positive species, the MMSO is divergent upstream of  dnaG . The MMSO of  Bacillus subtilis  contains three open reading frames (ORFs) that are differentially regulated by multiple promoters. In conjunction with studies to determine the expression profile of  dnaG , the MMSO of  Staphylococus epidermidis  was characterized.    Results  The ORFs of  S. epidermidis  were compared to the previously described MMSO of  B. subtilis  and two additional ORFs in  S. epidermidis, serp1129  and  serp1130 , were identified. The largest transcript, 4.8 kb in length, was expressed only in exponential growth and encompassed all four ORFs ( serp1130, serp1129, dnaG , and  sigA ). A separate transcript (1.5 kb) comprising  serp1130  and  serp1129  was expressed in early exponential growth. Two smaller transcripts 1.3 and 1.2 kb in size were detected with a  sigA probe  in both exponential and post-exponential phases of growth. Western blot analysis correlated with the transcriptional profile and demonstrated that Serp1129 was detected only in the exponential phase of growth. Computational analysis identified that Serp1130 contained a CBS motif whereas Serp1129 contained an ATP/GTP binding motif. Functional studies of Serp1129 demonstrated that it was capable of binding both ATP and GTP. Comparisons with a  sigB:dhfr  mutant revealed that the 1.3 kb  sigA  transcript was regulated by a σ B -dependent promoter.    Conclusions  These studies demonstrated that the  S. epidermidis  1457 MMSO contains two ORFs ( serp1129  and  serp1130 ) not described within the  B. subtilis  MMSO and at least three promoters, one of which is σ β -dependent. The transcriptional regulation of  sigA  by σ B  provides evidence that the staphylococcal σ B -dependent response is controlled at both the transcriptional and post-transcriptional level. The conservation of  serp1129  across multiple gram-positive organisms and its capability to bind ATP and GTP support the need for further investigation of its role in bacterial growth.

bmc microbiology

Genetic analysis of the Staphylococcus epidermidis Macromolecular Synthesis Operon: Serp1129 is an ATP binding protein and sigA transcription is regulated by both σA- and σB-dependent promoters