Escherichia coli cAMP receptor protein-DNA complexes. 1. Energetic contributions of half-sites and flanking sequences in DNA recognition

Biochemistry
E A PylesJ C Lee

Abstract

In Escherichia coli, the cyclic AMP receptor protein (CRP) serves as a sensor of the intracellular level of cyclic AMP. At increasing concentrations of cyclic AMP, CRP becomes activated upon binding a cyclic AMP molecule. The activated CRP is capable of regulating the expression of more than 20 genes by binding to specific DNA sites. The specific DNA sequences recognized by CRP consist of two-half-sites of the consensus sequence TGTGA......XCAXA. At present, the relative contributions of the two half-site and flanking sequences in the energetics of CRP recognition have not been quantitatively defined. A series of 20 DNA sequences was designed to dissect the contributions of individual half-sites and flanking sequences using the natural gal P1 and lac P1 sequences as the initial targets. The binding of CRP to these DNA sequences was monitored by fluorescence anisotropy. None of the individual half-sites or flanking sequences contributes more to the binding energetics than a random sequence. In the lac P1 sequence, the combination of both half-sites leads to a >100-fold increase in affinity compared to that of an individual half-site in CRP-DNA complex formation. The flanking sequence of lac P1 exhibits a 10- and 0-fold enhanceme...Continue Reading

References

Jan 1, 1993·Annual Review of Biochemistry·A KolbS Adhya

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Citations

May 10, 2001·Biochimica Et Biophysica Acta·J G Harman
Feb 20, 2010·Trends in Microbiology·Nicholas P TuckerMatthew I Hutchings

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