Factors Affecting Start Site Selection at the Escherichia coli fis Promoter

Abstract

ABSTRACT Transcription initiation with CTP is an uncommon feature among Escherichia coli σ 70 promoters. The fis promoter ( fis P), which is subject to growth phase-dependent regulation, is among the few that predominantly initiate transcription with CTP. Mutations in this promoter that cause a switch from utilization of CTP to either ATP or GTP as the initiation nucleotide drastically alter its growth phase regulation pattern, suggesting that the choice of the primary initiating nucleotide can significantly affect its regulation. To better understand what factors influence this choice in fis P, we made use of a series of promoter mutations that altered the nucleotide or position used for initiation. Examination of these promoters indicates that start site selection is determined by a combination of factors that include preference for a nucleotide distance from the −10 region (8 > 7 > 9 ≫ 6 ≫ 10 > 11), initiation nucleotide preference (A = G ≫ CTP ≥ UTP), the DNA sequence surrounding the initiation region, the position of the −35 region, and changes in the intracellular nucleoside triphosphate pools. We describe the effects that each of these factors has on start site selection in the fis P and discuss the interplay between position and nucleotide preference in this important process.