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Autogenous regulation is a general strategy of balancing ribosomal protein synthesis in bacteria. Control mechanisms have been studied in detail for most of ribosomal protein operons, except for  rpsB-tsf  encoding essential r-protein S2 and elongation factor Ts, where even the promoter has remained unknown. By using single-copy translational fusions with the chromosomal  lacZ  gene and Western-blot analysis, we demonstrate here that S2 serves as a negative regulator of both  rpsB  and  tsf  expression in vivo, acting at a single target within the  rpsB  5′-untranslated region (5′-UTR). As determined by primer extension, transcription of the  Escherichia coli rpsB-tsf  operon starts 162 nucleotides upstream of the  rpsB  initiation codon at a single promoter TGTGGTATAAA belonging to the extended −10 promoter class. Both the promoter signature and the 5′-UTR structure of the  rpsB  gene appear to be highly conserved in γ-proteobacteria. Deletion analysis of the  rpsB  5′-UTR within  rpsB′-′lacZ  fusions has revealed that an operator region involved in the S2 autoregulation comprises conserved structural elements located upstream of the  rpsB  ribosome binding site. The S2-mediated autogenous control is impaired in  rpsB  mutants and, more surprisingly, in the  rpsA  mutant producing decreased amounts of truncated r-protein S1 ( rpsA ∷IS 10 ), indicating that S2 might act as a repressor in cooperation with S1.

A new regulatory circuit in ribosomal protein operons: S2-mediated control of the rpsB-tsf expression in vivo