Detecting the DNA kinks in a DNA-CRP complex in solution with iodine-125 radioprobing.
Auger-electron-emitting radioisotopes such as 125I produce DNA strand breaks within nanometer range of the decay site. Here we analyze these breaks in order to study changes in DNA conformation upon binding with cyclic AMP receptor protein (CRP) in solution. The clear difference we found in break frequency in the CRP-DNA complex, as compared to the naked DNA duplex, correlates with the increased distances between the deoxyriboses and the radioiodine atom caused by the CRP-induced kink observed in the cocrystal. Thus, we demonstrate that 125I radioprobing can be used to study fine conformational changes of DNA within DNA-protein complexes.
Karamychev
V N
VN
Nuclear Medicine Department, National Institutes of Health, Bethesda, Maryland 20892-1180, USA.
Zhurkin
V B
VB
Garges
S
S
Neumann
R D
RD
Panyutin
I G
IG
D001483:Base Sequence; D002373:Cyclic AMP Receptor Protein / Q000737:chemistry*; D004247:DNA / Q000737:chemistry*; D015342:DNA Probes; D007457:Iodine Radioisotopes; D008958:Models, Molecular; D009690:Nucleic Acid Conformation
D002373:Cyclic AMP Receptor Protein; D015342:DNA Probes; D007457:Iodine Radioisotopes; D004247:DNA