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Micrococcal nuclease as a DNA structural probe: Its recognition sequences, their genomic distribution and correlation with DNA structure determinants

1986; Elsevier BV; Volume: 190; Issue: 4 Linguagem: Inglês

10.1016/0022-2836(86)90247-0

1089-8638

James T. Flick, Joel C. Eissenberg, Sarah C. R. Elgin,

Enterobacteriaceae and Cronobacter Research

We have analyzed micrococcal nuclease (MNase) DNA cleavage patterns at the sequence level by examining 2.3 × 103 base-pairs of data derived from the Drosophila melanogaster 44D larval cuticle locus. Within this region, MNase preferentially cleaved 140 sites. Clusters of these sites appear to generate the preferential MNase eukaryotic DNA cleavage sites seen on agarose gels at roughly 100 to 300 base-pair intervals. These clusters of preferential cleavage sites rarely occur within gene coding regions. The analysis revealed that duplex DNA sequences preferentially cleaved by MNase are generally determined by a single strand sequence: d(A-T)n, where n ⩾ 1, flanked by a 5′ dC or dG. Cleavage of the other strand is generally staggered 5′ by several nucleotides and occurs even if such sequences are absent on that strand. An empirical predictive DNA cleavage model derived from a statistical analysis of the sequence level data was applied to seven eukaryotic gene loci of known sequence. The predicted patterns were in good general agreement with the previously observed eukaryotic gene/spacer cleavage pattern. Statistical analysis also revealed that sites of predicted preferential DNA cleavage occur less frequently in protein coding regions than for randomized sequences of the same length and nucleotide content. Comparison of the MNase cleavage patterns to the sequence-dependent pattern of binding energies between duplex DNA strands indicates that MNase preferentially cleaves sequences with low helix stability.