Expression of fnr Is Constrained by an Upstream IS 5 Insertion in Certain Escherichia coli K-12 Strains

Abstract

ABSTRACT FNR is a global transcriptional regulator that controls anaerobic gene expression in Escherichia coli . Through the use of a number of approaches it was shown that fnr gene expression is reduced approximately three- to fourfold in E. coli strain MC4100 compared with the results seen with strain MG1655. This reduction in fnr expression is due to the insertion of IS 5 ( is5F ) in the regulatory region of the gene at position −41 relative to the transcription initiation site. Transcription of the fnr gene nevertheless occurs from its own promoter in strain MC4100, but transcript levels are reduced approximately fourfold compared with those seen with strain MG1655. Remarkably, in strains bearing is5F the presence of Hfq prevents IS 5 -dependent transcriptional silencing of fnr expression. Thus, an hfq mutant of MC4100 is devoid of FNR protein and has the phenotype of an fnr mutant. In strain MG1655, or a derivative of MC4100 lacking is5F , mutation of hfq had no effect on fnr transcript levels. This finding indicates that IS 5 mediates the effect of Hfq on fnr expression in MC4100. Western blot analysis revealed that cellular levels of FNR were reduced threefold in strain MC4100 compared with strain MG1655 results. A selection of FNR-dependent genes fused to lacZ were analyzed for the effects of reduced FNR levels on anaerobic gene expression. Expression of some operons, e.g., focA - pfl and fdnGHJI , was unaffected by reduction in the level of FNR, while the expression of other genes such as ndh and nikA was clearly affected.