Transcription products from the rplKAJL- rpoBC gene cluster

Abstract

Transcripts from the rplKAJL- rpoBC ribosomal protein—RNA polymerase gene cluster have been quantified and their ends mapped using RNA—DNA hybridization, sucrose density-gradient sedimentation, Northern hybridization and S 1 nuclease protection. The results indicate that the most abundant transcript is the 2600 nucleotide tetracistronic L11-L1-L10-L12 mRNA initiated at the upstream major P L11 promoter and terminated at the transcription attenuator in the L12-β intergenic space. Somewhat less abundant 1300 nucleotide L11-L1 and L10-L12 bicistronic transcripts were observed. The 3′ ends of the L11-L1 transcripts were heterogeneous; most of the ends were localized to three sites within a 110 base-pair region in the L1-L10 intergenic space. This intergenic space encodes also the major P L10 promoter and the mRNA binding site for the L10 translational control protein. Two 5′ ends were observed for L10-L12 bicistronic mRNA, one at the P L10 promoter and the other 150 nucleotides further downstream in a region in which promoter activity has not been detected. It is suggested that this second downstream 5′ end is generated by processing of the transcripts initiated at the major P L10 promoter. No transcript initiation in the L10-L12 intergenic space was detected. About 80% of the transcripts reading through the L12 gene were terminated in the vicinity of the transcription attenuator that is responsible for the reduction in the expression of the downstream RNA polymerase genes. Transcripts reading through the attenuator were partially processed by RNase III within a potential hairpin structure in the RNA transcript. Processing appears to produce 3′ and 5′ transcript end sites separated by about ten nucleotides. No other major 5′ ends were observed in the L12-β intergenic space. These results indicate that the two major promoters, P L11 and P L10, are both utilized to drive the interrelated transcriptional expression of this ribosomal protein-RNA polymerase gene cluster.