CRP-DNA complexes: inducing the A-like form in the binding sites with an extended central spacer.
The consensus DNA sequence for binding of the Escherichia coli cyclic AMP receptor protein (CRP) has two symmetrically related inverted recognition elements TGTGA:TCACA, separated by a variable spacer, normally 6 bp long. We have shown that the CRP-cAMP complex, when bound to synthetic binding sites with an extended 8 bp spacer segment, induces an increase in the DNA circular dichroism (CD). The CD change at lambda > 275 nm agrees with the shift of approximately one helical turn of DNA into A-like form. The B-conformation is preserved for CRP binding sites similar to that in the lac and uxaCA promoters with 6 bp spacers. Another effect accompanying DNA binding is a dramatic increase of the negative CD magnitude in the spectral region of the ligand cAMP, at lambda < 272 nm. This effect is observed when CRP binds to specific sites with 6 or 8 bp spacers as well as to non-specific DNA. We reason that the A-like form arises by compressing and unwinding the DNA in CRP-DNA complexes having 8 bp central spacers. This serves to maintain a fixed length and twisting angle and is controlled by the protein's relatively rigid frame. This model is consistent with the observation that some binding sites with 6 bp spacers may also show the CD increase inherent to the sites with the extended 8 bp spacers. These 6 bp spacers are characterized by an increased twisting angle that requires their unwinding to bind to CRP. We propose that a mutual adaptation between CRP and binding sites by local untwisting and a B-->A-like transition in the DNA is of general importance and may occur in other protein-DNA complexes, such as the complex of RNA polymerase with promoter DNA.
National Cancer Institute, National Institutes of Health, Bethesda, MD 20892.
V I
VI
Ivanov
L E
LE
Minchenkova
B K
BK
Chernov
P
P
McPhie
S
S
Ryu
S
S
Garges
A M
AM
Barber
V B
VB
Zhurkin
S
S
Adhya
D001483:Base Sequence; D002942:Circular Dichroism; D000242:Cyclic AMP / Q000737:chemistry / Q000378:metabolism; D002373:Cyclic AMP Receptor Protein / Q000378:metabolism*; D004247:DNA / Q000737:chemistry* / Q000378:metabolism*; D004269:DNA, Bacterial / Q000737:chemistry / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D011485:Protein Binding
D002373:Cyclic AMP Receptor Protein; D004269:DNA, Bacterial; D004247:DNA; D000242:Cyclic AMP