Activation of Escherichia coli leuV transcription by FIS.
The transcription factor FIS has been implicated in the regulation of several stable RNA promoters, including that for the major tRNALeu species in Escherichia coli, tRNA1Leu. However, no evidence for direct involvement of FIS in tRNA1Leu expression has been reported. We show here that FIS binds to a site upstream of the leuV promoter (centered at -71) and that it directly stimulates leuV transcription in vitro. A mutation in the FIS binding site reduces transcription from a leuV promoter in strains containing FIS but has no effect on transcription in strains lacking FIS, indicating that FIS contributes to leuV expression in vivo. We also find that RNA polymerase forms an unusual heparin-sensitive complex with the leuV promoter, having a downstream protection boundary of approximately -7, and that the first two nucleotides of the transcript, GTP and UTP, are required for formation of a heparin-stable complex that extends downstream of the transcription start site. These studies have implications for the regulation of leuV transcription.
Ross
W
W
Department of Bacteriology, University of Wisconsin, Madison, Wisconsin 53706, USA.
Salomon
J
J
Holmes
W M
WM
Gourse
R L
RL
D001483:Base Sequence; D002352:Carrier Proteins / Q000378:metabolism*; D018983:DNA Footprinting; D004268:DNA-Binding Proteins / Q000378:metabolism; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D003850:Deoxyribonuclease I; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D029968:Escherichia coli Proteins; D035021:Factor For Inversion Stimulation Protein; D015964:Gene Expression Regulation, Bacterial; D005838:Genotype; D034982:Integration Host Factors; D007700:Kinetics; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D012356:RNA, Transfer, Leu / Q000235:genetics*; D014158:Transcription, Genetic
D002352:Carrier Proteins; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; D035021:Factor For Inversion Stimulation Protein; D034982:Integration Host Factors; D012356:RNA, Transfer, Leu; C440502:integration host factor, E coli; D012321:DNA-Directed RNA Polymerases; D003850:Deoxyribonuclease I