Design of cAMP-CRP-activated promoters in Escherichia coli.
We have studied the deoP2 promoter of Escherichia coli to define features that are required for optimal activation by the complex of adenosine 3',5' monophosphate (cAMP) and the cAMP receptor protein (CRP). Systematic mutagenesis of deoP2 shows that the distance between the CRP site and the -10 hexamer is the crucial factor in determining whether the promoter is activated by cAMP-CRP. Based on these observations, we propose that cAMP-CRP-activated promoters can be created by correctly aligning a CRP target and a -10 hexamer. This idea has been successfully tested by converting both a CRP-independent promoter and a sequence resembling the consensus -10 hexamer to strongly cAMP-CRP-activated promoters.
Valentin-Hansen
P
P
Department of Molecular Biology, University of Odense, Denmark.
Holst
B
B
Søgaard-Andersen
L
L
Martinussen
J
J
Nesvera
J
J
Douthwaite
S R
SR
D001483:Base Sequence; D000242:Cyclic AMP / Q000378:metabolism*; D004269:DNA, Bacterial; D004926:Escherichia coli / Q000235:genetics*; D015964:Gene Expression Regulation, Bacterial; D008969:Molecular Sequence Data; D016296:Mutagenesis; D011401:Promoter Regions, Genetic; D011953:Receptors, Cyclic AMP / Q000378:metabolism*
D004269:DNA, Bacterial; D011953:Receptors, Cyclic AMP; D000242:Cyclic AMP