Bacillus subtilis pur operon expression and regulation

Abstract

The Bacillus subtilis pur operon is a 12-gene cluster, purEKB-purC(orf)QLF-purMNH(J)-purD, organized in groups of overlapping coding units separated by intercistronic gaps. Translational fusions of Escherichia coli lacZ were constructed to purE, purC, and purM, the first gene of each group. Analyses of gene fusions integrated into the chromosomal pur operon exclude the possibility of internal promoters in intercistronic regions and support the view that transcription is from the single sigma 43 promoter at the 5' end of the operon. Enzyme and mRNA measurements indicate that transcriptional regulation occurs solely at the 5' end of the operon. The relative levels of beta-galactosidase from purE-lacZ, purC-lacZ, and purM-lacZ were determined under repressing and nonrepressing conditions. These results indicate that expression of purC-lacZ was 3.0- to 6.8-fold higher than purE-lacZ because of enhanced translational efficiency. The enhanced translational efficiency of purC-lacZ was accompanied by a partial escape from regulation by purines. This anomalous effect on purC-lacZ was the only suggestion for posttranscriptional regulation.