A sigma 54 promoter and downstream sequence elements ftr2 and ftr3 are required for regulated expression of divergent transcription units flaN and flbG in Caulobacter crescentus.

Abstract

In this study, we investigated the cis-acting sequences required for transcription of the divergent, cell cycle-regulated flaN and flbG operons of Caulobacter crescentus. Previous work showed that transcription of flbG in vivo depends on a sigma 54 promoter and a sequence element called ftr1 that is located about 100 bp upstream from the transcription start site (D. A. Mullin and A. Newton, J. Bacteriol. 171:3218-3227, 1989). We now show that regulation of flaN transcription in vivo depends on a sigma 54 promoter and two ftr elements located downstream of the transcription start site at +86 (ftr2) and +120 (ftr3). Mutations in or between the conserved elements at -24 and -12 in this sigma 54 promoter reduced or abolished flaN transcription, and one mutation that eliminated flaN expression led to an increased level of flbG transcript. Mutations in ftr2 resulted in greatly reduced levels of flaN transcript but had no noticeable effect on flbG transcript levels. All three mutations constructed in ftr3 resulted in elevated flaN and flbG transcript levels. We conclude that ftr2 is required for positive regulation of flaN, whereas ftr3 appears to play a negative regulatory role in flaN and flbG expression. To explain the coordinated positive activation and negative autoregulation of these two transcription units and the effect of mutations on gene expression, we propose a model in which the flaN and flbG promoters interact through alternative DNA looping to form structures that are transcriptionally active or inactive.