CHARACTERIZATION OF A PROTEIN-BINDING SEQUENCE IN THE PROMOTER REGION OF THE 16S RIBOSOMAL-RNA GENE OF THE SPINACH CHLOROPLAST GENOME

By means of mobility-shift assays and Exonuclease III mapping we have determined a 14 bp sequence (named CDF2 binding site) located in front of the 16S rRNA initiation start site which is protected by a spinach chloroplast extract. This region does not include neither one of the two '-35' nor of the two '-10' E. coli-like promoter elements which are recognised by E. coli RNA polymerase in vitro. The CDF2 binding site is specifically recognized by two small polypeptides which migrate corresponding to 35 and 33 kDa respectively as shown by UV cross-linking experiments. In vivo transcription initiation of the 16S rRNA gene occurs 13 nucleotides downstream of the 14 bp sequence and is different from the transcription start site which is used by E. coli polymerase in vitro.