STRUCTURE OF THE 5' UPSTREAM REGION AND THE REGULATION OF THE RPOS GENE OF ESCHERICHIA-COLI

The nucleotide sequence of the 5' upstream region of the Escherichia c oli rpoS gene was determined and analyzed. At least four promoters res ponsible for rpoS transcription were identified, and designated PI, P2 , P3 and P4, P1 being furthest from the upstream. Using lacZ fusion ge nes, the P2 promoter was found to be the strongest of the four. All of these promoters are regulated similarly, and their activity is enhanc ed 2 to 3-fold in stationary phase. P1 and P2 transcription start site s were determined by primer extension analyses. The P2 promoter region shows similarity to the consensus sigma(70)-type promoter sequence, a nd was recognized by a both E sigma(70) and E sigma(38) holoenzymes in vitro. The mRNA transcribed from the most distal promoter, P1, appear s to include another open reading frame (orf-281), indicating that the two open reading frames comprise an operon. The rpoS gene product (si gma(38)) was rapidly degraded after addition of chloramphenicol to cul tures in the exponential, but not the stationary phase. This strongly suggests that posttranslational regulation is involved in the control of rpoS expression.