Direct evidence for autogenous regulation of the Escherichia coli genes rpoBC in vivo

We have fused the rpoBC genes to the strong controllable promoter PL in phage lambda while deleting most of the intercistronic regulatory DNA and ribosomal protein genes upstream of rpoB. Induction of a lysogen carrying the recombinant prophage gave rise to a 2-3-fold oversynthesis of beta beta' in the cell whereas rpoBC-mRNA levels rose by at least 10-fold. Similar observations were made when these sequences were present in the prophage, indicating that the removal of DNA sequences up to 26 base pairs before rpoB does not affect post-transcriptional autogenous regulation of beta beta' synthesis. Overexpression of beta beta' also autogenously regulated the synthesis of the beta polypeptide from the chromosome in two strains carrying electrophoretic mobility mutations in rpoB. S1 nuclease mapping experiments indicated that this regulation was also post-transcriptional, and confirmed that phage beta-mRNA synthesis exceeded chromosomal beta-mRNA synthesis by 20-fold. The provision of excess beta alone in the cell caused autoregulation of chromosomal beta, but not beta' synthesis, indicating that beta and beta' are regulated independently.