The role of FIS in trans activation of stable RNA operons of E. coli.
The thrU(tufB) operon of Escherichia coli is endowed with a cis-acting region upstream of the promoter, designated UAS for Upstream Activator Sequence. A protein fraction has been isolated that binds specifically to DNA fragments of the UAS, thus forming three protein-DNA complexes corresponding to three binding sites on the UAS. It stimulates in vitro transcription of the operon by facilitating the binding of the RNA polymerase to the promoter. All three protein-DNA complexes contain one and the same protein. Dissociation constants for the three complexes have been determined, the lowest being in the sub-nanomolar range. The protein also binds to the UAS of the tyrT operon and to the UAS upstream of the P1 promoter of the rrnB operon, suggesting that transcription of the three operons, if not of more stable RNA operons, is activated by a common trans activator. We demonstrate that the E.coli protein FIS (Factor for Inversion Stimulation) also binds to the UAS of the thrU(tufB) operon forming three protein-DNA complexes. A burst of UAS- and FIS-dependent promoter activity is observed after reinitiation of growth of stationary cultures in fresh medium.
Department of Biochemistry, Leiden University, Gorlaeus Laboratories, The Netherlands.
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L
Nilsson
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Vanet
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Vijgenboom
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Bosch
D002352:Carrier Proteins / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics*; D029968:Escherichia coli Proteins; D035021:Factor For Inversion Stimulation Protein; D006153:Guanosine Diphosphate / Q000378:metabolism; D034982:Integration Host Factors; D007700:Kinetics; D009876:Operon; D010444:Peptide Elongation Factor Tu / Q000378:metabolism; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D012329:RNA, Bacterial / Q000235:genetics*; D014158:Transcription, Genetic; D015533:Transcriptional Activation
D002352:Carrier Proteins; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; D035021:Factor For Inversion Stimulation Protein; D034982:Integration Host Factors; D012329:RNA, Bacterial; C440502:integration host factor, E coli; D006153:Guanosine Diphosphate; D010444:Peptide Elongation Factor Tu