Positive regulation of the colicin E1 gene by cyclic AMP and cyclic AMP receptor protein

In previous experiments, we showed that the in vivo transcription of the colicin E1 gene was dependent on cyclic AMP in adenylate cyclase-defective mutant cells of Escherichia coli (Ebina, Y. and Nakazawa, A (1983) J. Biol. Chem. 258, 7072-7078). We now show that cyclic AMP and cyclic AMP receptor protein stimulated the in vitro transcription of the gene in the presence of spermidine. As determined in DNase I protection experiments, two binding sites for the complex of cyclic AMP and the receptor protein were identified about 60 base pairs (CRP-1) and 110 base pairs (CRP-2) upstream from the transcription initiation site of the colicin E1 gene. CRP-1 had a higher affinity for the complex than that of CRP-2. Substituting an unrelated DNA sequence for CRP-2 reduced the efficiency of in vitro stimulation of the gene by cyclic AMP and the receptor protein. These potential binding sites for the cyclic AMP-cyclic AMP receptor protein complex probably participate in the stimulation of the colicin E1 gene transcription.