Investigations on stimulation of lac transcription in vivo in Escherichia coli by cAMP analogues

Abstract

The ability of 24 systematically modified analogues of adenosine 3′,5′‐monophosphate (cAMP) to enhance the synthesis of β‐galactosidase in glucose‐repressed Escherichia coli strains KNBL 1001 and cpd – Crookes has been investigated. The properties of the analogues in comparison with cAMP are, with only two exceptions, alike in both strains. Two analogues, 7‐deazaadenosine 3′,5′‐monophosphate (i.e. tubercidin 3′,5′‐monophosphate) and ( R p )‐adenosine 3′,5′‐monothionophosphate, exhibit higher biological activity than cAMP. The latter analogue is 50‐fold more active in both strains. Three analogues showed activities comparable to cAMP, four analogues were less active and 12 analogues were unable to antagonize catabolite repression. Structure‐activity correlations showed that the 2′OH‐, 3′O‐, 5′O‐, the negative charge and the 6‐amino group cannot be modified without losing biological activity in vivo , while the N‐1 and N‐7 in adenine are not essential. The interaction with the catabolite gene activator protein is stereoselective for an unmodified axial exocyclic oxygen. The results are compared to those obtained with cAMP analogues in E. coli in vitro and those obtained with the same analogues in protein‐kinase systems and Dictyostelium species. The model of McKay et al. [McKay, D. B., Weber, J. T. and Steitz, T. A. (1982) J. Biol. Chem. 257 , 9518–9524] proposed for distinct chemical interactions of cAMP with the catabolite gene activator protein is discussed and supplemented by additional hydrogen bond interactions.