J. Biol. Chem., Vol. 261, Issue 22, 10051-10057, Aug, 1986
UV-induced interstrand cross-linking of d(GT)n.d(CA)n is facilitated by a structural transition
JD Love, HT Nguyen, A Or, AK Attri and KW Minton
Photochemical alterations following ultraviolet irradiation of the
alternating copolymer d(GT)n.d(CA)n were studied. We found that in solution
conditions which produced circular dichroism spectra compatible with B-form
or A-form DNA, no interstrand cross-linking or photoproduct formation could
be demonstrated. Zimmer et al. (Zimmer, C., Tymen, S., Marck, C., and
Guschlbaumer, W. (1982) Nucleic Acids Res. 10, 1081-1091) and Vorlickova et
al. (Vorlickova, M., Kypr, J., Sotkrova, S., Sponar, J. (1982) Nucleic
Acids Res. 10, 1071-1080) have reported a number of solution conditions
which produce a structural transition of this polymer characterized by a
negative deviation of the circular dichroism spectrum in the region of 280
nm. The nature of this transition has not yet been elucidated. Following
ultraviolet irradiation of d(GT)n.d(CA)n under two conditions which produce
this transition (manganese solution or ethanol plus trace salts solution)
we found ultraviolet dose-dependent interstrand cross-linking as well as
dose-dependent formation of thymine-containing photoproduct. Interstrand
cross-linking is demonstrated by two criteria: increase in polymer size as
detected by alkaline agarose gel electrophoresis, and generation of
intermediate density material in alkaline cesium sulfate isopycnic
gradients. The thymine-containing photo-product was demonstrated by thin
layer chromatography of acid hydrolysates of the polymer. The photo-product
is at least partially photoreversible. These findings suggest that the
geometry of the alternative conformation is such that pyrimidines from
different strands are closely approximated, allowing for photodimerization.