UTP Allosterically Regulates Transcription by Escherichia coli RNA Polymerase from the Bacteriophage T7 A1 Promoter

• Published in: Journal of molecular biology 2002-04, Vol.318 (2), p.305-320
• Main Authors: Johnson, Ronald S, Chester, Rebecca E
• Format: Article
• Language: English
• Subjects: Adenosine Triphosphate - metabolism; Allosteric Regulation; Bacteriophage T7 - genetics; DNA-Directed RNA Polymerases - metabolism; Escherichia coli - drug effects; Escherichia coli - genetics; Escherichia coli - metabolism; Escherichia coli RNA polymerase; Kinetics; Oligoribonucleotides - metabolism; promoter clearance; Promoter Regions, Genetic; Spectrometry, Fluorescence; transcription initiation; Transcription, Genetic - drug effects; transcriptional pausing; Uridine Triphosphate - metabolism; Uridine Triphosphate - pharmacology
• ISSN: 0022-2836; 1089-8638
• DOI: 10.1016/S0022-2836(02)00042-6

In the case of Escherichia coli RNA polymerase, UTP at elevated concentrations suppresses terminated transcript accumulation during multiple-round transcription from a DNA construct containing the T7 A1 promoter and T e terminator. The step that is affected by UTP at elevated concentrations is promoter clearance. In an attempt to understand better the mechanism by which UTP regulates this step, we analyzed the effect of UTP on the formation of pppApU in the presence of only UTP and ATP. At elevated concentrations, UTP is a non-competitive inhibitor with respect to ATP in the formation of pppApU. This indicates that the effect of UTP on the formation of pppApU is mediated through an allosteric site. Moreover, the magnitude of the inhibition of pppApU formation is sufficient to account for the decrease in terminated transcript accumulation at elevated UTP concentrations. Thus, it appears that UTP modulates terminated transcript accumulation during multiple-round transcription from this DNA construct by allosteric regulation of promoter clearance at the point of transcription initiation.