RNA polymerase active centre compensates for the absence of transcription proofreading factors in Cyanobacteria

Abstract

SummaryThe vast majority of organisms possess transcription elongation factors, the functionally similar bacterial Gre and eukaryotic TFIIS/TFS. Their main cellular functions are to proofread errors of transcription and to restart elongation via stimulation of RNA hydrolysis by the active centre of RNA polymerase (RNAP). Very few taxons lack these factors, including the large evolutionarily ancient group of cyanobacteria and their descendants, the chloroplasts. How do they cope? What compensatory mechanisms they possess?We found that cyanobacterial RNAP functionally substitutes for Gre/TFIIS - it does not stall on DNA, it efficiently catalyses the proofreading reaction of RNA hydrolysis, and the drop in transcription fidelity is only fractional, as confirmed by NGS. This alternative, presumably primordial, route to fidelity and processivity in the absence of Gre/TFIIS factors is based on the active site of RNAP stabilisation in a closed conformation. However, here lies a trade off - a severely reduced ability of this active site to recognise regulatory pausing signals. We suggest that perhaps the main advantage of Gre/TFIIS acquisition was to allow transcription regulation via pausing; with increase in fidelity as a bonus side effect.