The Escherichia coli Fis promoter is regulated by changes in the levels of its transcription initiation nucleotide CTP.
Expression of the Escherichia coli nucleoid-associated protein Fis (factor for inversion stimulation) is controlled at the transcriptional level in accordance with the nutritional availability. It is highly expressed during early logarithmic growth phase in cells growing in rich medium but poorly expressed in late logarithmic and stationary phase. However, fis mRNA expression is prolonged at high levels throughout the logarithmic and early stationary phase when the preferred transcription initiation site (+1C) is replaced with A or G, indicating that initiation with CTP is a required component of the regulation pattern. We show that RNA polymerase-fis promoter complexes are short lived and that transcription is stimulated over 20-fold from linear or supercoiled DNA if CTP is present during formation of initiation complexes, which serves to stabilize these complexes. Use of fis promoter fusions to lacZ indicated that fis promoter transcription is sensitive to the intracellular pool of the predominant initiating NTP. Growth conditions resulting in increases in CTP pools also result in corresponding increases in fis mRNA levels. Measurements of NTP pools performed throughout the growth of the bacterial culture in rich medium revealed a dramatic increase in all four NTP levels during the transition from stationary to logarithmic growth phase, followed by reproducible oscillations in their levels during logarithmic growth, which later decrease during the transition from logarithmic to stationary phase. In particular, CTP pools fluctuate in a manner consistent with a role in regulating fis expression. These observations support a model whereby fis expression is subject to regulation by the availability of its initiating NTP.
Walker
Kimberly A
KA
Department of Biological Sciences, University at Albany, Albany, New York 12222, USA.
Mallik
Prabhat
P
Pratt
Timothy S
TS
Osuna
Robert
R
D001483:Base Sequence; D015152:Blotting, Northern; D002855:Chromatography, Thin Layer; D003562:Cytidine / Q000737:chemistry; D003570:Cytidine Triphosphate / Q000737:chemistry*; D017931:DNA Primers / Q000737:chemistry; D004278:DNA, Superhelical / Q000737:chemistry / Q000235:genetics; D012321:DNA-Directed RNA Polymerases / Q000737:chemistry / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D035021:Factor For Inversion Stimulation Protein / Q000235:genetics*; D015971:Gene Expression Regulation, Enzymologic; D007700:Kinetics; D007763:Lac Operon; D008954:Models, Biological; D008957:Models, Genetic; D008969:Molecular Sequence Data; D009991:Oscillometry; D010957:Plasmids / Q000378:metabolism; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000378:metabolism; D012492:Salts / Q000494:pharmacology; D013997:Time Factors; D014158:Transcription, Genetic; D001616:beta-Galactosidase / Q000378:metabolism
D017931:DNA Primers; D004278:DNA, Superhelical; D035021:Factor For Inversion Stimulation Protein; D012333:RNA, Messenger; D012492:Salts; D003562:Cytidine; D003570:Cytidine Triphosphate; D012321:DNA-Directed RNA Polymerases; D001616:beta-Galactosidase