Detection of an unusual distortion in A-tract DNA using KMnO4: effect of temperature and distamycin on the altered conformation.
The chemical probes potassium permanganate (KMnO4) and diethylpyrocarbonate (DEPC) can be used to study the conformational flexibility of short tracts of adenine (A-tracts) present in DNA. With these probes, we demonstrate that a novel distortion is induced in a 5 base pair A-tract at low temperature. Formation of this distorted A-tract structure, which occurs in a DNA fragment from the promoter region of the plasmid pBR322, is distinguished by a dramatic increase in the KMnO4 reactivity of the central thymines in this tract at 12 degrees C. This alteration occurs in the absence of any detectable rearrangement in the conformation of the adenines in the complementary strand. Induction of this low temperature A-tract structure is blocked by the minor groove binding drug distamycin. Hydroxyl radical footprinting of distamycin binding to the fragment containing the d(A)5 tract at 12 degrees C suggests that this drug has two different modes of binding to DNA in agreement with recent NMR data. These experiments show that short A-tracts are capable of forming more than one structural variant of B DNA in solution. The possible relationship between the intrinsic bending of DNA containing short phased A-tracts and the low temperature A-tract conformation is discussed.
McCarthy
J G
JG
Department of Biology, Massachusetts Institute of Technology, Cambridge 02139.
Rich
A
A
D000225:Adenine / Q000737:chemistry*; D001483:Base Sequence; D004247:DNA / Q000737:chemistry*; D004214:Distamycins / Q000494:pharmacology*; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation / Q000187:drug effects; D010957:Plasmids; D011071:Poly T / Q000737:chemistry; D011196:Potassium Permanganate / Q000494:pharmacology*; D011401:Promoter Regions, Genetic; D015183:Restriction Mapping; D013696:Temperature; D013816:Thermodynamics
D004214:Distamycins; D011196:Potassium Permanganate; D011071:Poly T; D004247:DNA; D000225:Adenine