A highly homogeneous 145-base-pair fragment of double helical poly(dA-dT) . poly(dA-dT) was obtained by micrococcal nuclease digestion of a semisynthetic chromatin prepared from the nucleosome core histones (H2A, H2B, H3, H4) and the synthetic polydeoxyribonucleotide. In contrast to higher molecular weight alternating copolymers, this fragment displayed two resolved 31P NMR signals, separated by 24 Hz at 10.93 MHz. The two signals were of equal intensity at all temperatures less than the Tm for the fragment. Analyses of the possible origins for the two reasonances leads to the conclusion that the phosphodiester backbone of this DNA contains two distinct phosphorus environments, probably in an alternating array. We suggest that this may indicate the presence of sequence-dependent local variation in the helical structure of DNA in general.
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