The binding of the cyclic AMP receptor protein to synthetic DNA sites containing permutations in the consensus sequence TGTGA
- 15 August 1987
- journal article
- research article
- Published by Portland Press Ltd. in Biochemical Journal
- Vol. 246 (1) , 227-232
- https://doi.org/10.1042/bj2460227
Abstract
The binding of the cyclic AMP receptor protein (CRP) to symmetrical synthetic DNA-binding sites was investigated with a gel-retardation assay. A set of ten different sequences was employed, comprising all base permutations at positions 2, 4, and 5 of the consensus sequence 5′(TGTGA)3′. We show that: (i) CRP has a higher affinity for the completely symmetrical site than towards the lac wild-type site; (ii) base substitutions at position 2 lead to either a complete loss of specific CRP binding (G—C), a reduction in specific CRP binding (G—A) or only marginal effects on specific CRP binding (G—T); (iii) changes at position 4 abolish (G—C; G—A) or reduce (G—T) specific CRP binding; and (iv) base permutations at position 5 reduce specific CRP binding, but never completely abolish it. Thus position 4, and to a lesser extent position 2, in the DNA consensus sequence are the most crucial ones for specific binding by CRP.This publication has 17 references indexed in Scilit:
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