Analysis of chromatin structure and DNA sequence organization: use of the 1,10-phenanthroline-cuprous complex.
Limited treatment of Drosophila nuclei with the 1,10-phenanthroline-cuprous complex leads to rapid production of nucleosomal ladders indistinguishable from those obtained by micrococcal nuclease digestion. An investigation of the preferential sites of cleavage of protein-free DNA at locus 67B1 surprisingly indicated that both reagents recognized very similar features. Thus, a virtually identical pattern of preferential cleavages was generated over a 12 kb fragment encoding four transcripts at this locus. The distribution of cleavage sites was highly non-random, with major sites falling in the spacers between the genes. Both reagents cleaved certain chromatin-specific sites near the 5' ends of the genes. However, an analysis of preferential cleavages at the sequence level did not reveal the same close correspondence. We suggest that both reagents can recognize some localized secondary structural features of the DNA and that the particular distribution of sequences present at this locus results in a distinctive pattern of cleavage sites that delineates gene and spacer segments.
D000818:Animals; D001483:Base Sequence; D002843:Chromatin / Q000032:analysis*; D004247:DNA / Q000235:genetics*; D004262:DNA Restriction Enzymes; D004330:Drosophila / Q000235:genetics; D008836:Micrococcal Nuclease; D009707:Nucleosomes / Q000032:analysis; D010618:Phenanthrolines; D010957:Plasmids
D002843:Chromatin; D009707:Nucleosomes; D010618:Phenanthrolines; C014741:bis(1,10-phenanthroline)copper(2+) ion; D004247:DNA; D004262:DNA Restriction Enzymes; D008836:Micrococcal Nuclease