Two promoters within the psbK-psbI-trnG gene cluster in tobacco chloroplast DNA

Abstract

Transcription of the 2.6 kbp psbK-psbI-trnG cluster in tobacco chloroplasts has been studied. This cluster contains, in linear sequence, the genes encoding two low-molecular-mass polypeptides, K and I, of photosystem II (psbK and psbI, respectively), and tRNA(Gly) (UCC) (trnG). Northern blot hybridization revealed that the largest transcript (2.6 kb) hybridizes to psbK, psbI and trnG, but not to the following trnR-UCU. Ten other transcripts ranging from 0.1 to 1.3 kb were also detected. Three of these transcripts overlap the divergent transcript arising from trnS-GCU located on the opposite DNA strand. S1 mapping and primer extension experiments showed that these multiple transcripts comprise eight distinct 5' ends. By in vitro capping assays two of them were determined to be transcriptional initiation sites; one is located 163 bp upstream of psbK and the other is 6 bp upstream of trnG. The 3' ends of transcripts were determined by S1 mapping; one lies between psbI and trnG and the other is at the end of trnG. The presence of dual promoters of trnG is discussed.