Abstract
The control of transcription initiation at the lactose operon promoter was investigated in vitro. We found that an upstream promoter (termed lac P 2) interfered with RNA polymerase binding at the principal promoter (termed lac P 1). The start site for lac P 2 was located at base pair position −22 relative to the P 1 start site. The addition of cAMP receptor protein and cAMP was shown to repress lac P 2 and to activate lac P 1. Abortive initiation reactions for both promoters were used to investigate the coordinate repression-activation elicited by CRP-cAMP. The effects of lac promoter mutations (L8, P 8, and UV5) were consistent with an important RNA polymerase positioning role for CRP-cAMP in the activation of lac operon expression.
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