Location of the tufB promoter of E. coil: Cotranscription of tufB with four transfer RNA genes

Abstract

Previous nucleotide sequence studies have demonstrated that the structural genes for four transfer RNA species ( thrU, tyrU, glyT and thrT) are positioned close to one another and near tufB, one of the structural genes for elongation factor Tu. We have carried out experiments to determine the position of the tufB promoter and thus to infer whether these five genes are in a single transcription unit. tufB cloned on plasmids was fused to Tc (in operon fusion) or to lacZ (in gene fusion). These plasmids were then subjected to in vitro deletion to locate the promoter responsible for tufB transcription. In addition, the ability of wild-type tufB to complement kirromycin resistance was determined with deletion plasmids. The results indicate that the major promoter for tufB lies upstream from the four transfer RNA genes, and that there might be at least one weak internal promoter, possibly adjacent to tufB. Assuming that the four tRNA genes that lie between the major promoter and tufB are also transcribed from that promoter, we suggest that all five genes lie in a single transcription unit ( thrUp-thrU-tyrU-glyT-thrT-tufB-tufBt) whose primary transcript is thus both a transfer RNA precursor and a messenger RNA.