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The chromatin structure of specific genes part 1 evidence for higher order domains of defined dna sequence






Cell 16(4): 797-806

The chromatin structure of specific genes part 1 evidence for higher order domains of defined dna sequence

When the chromatin of Drosophila melanogaster is examined by digestion with DNAse I or micrococcal nuclease, no general structural organization above the level of the nucleosome is revealed by the cleavage pattern. The DNAse I cleavage pattern of specific regions of the Drosophila chromosome shows discrete bands with sizes ranging from a few kilobase pairs (kb) to more than 20 kb. Visualization of such higher order bands was achieved by the use of the Southern blotting technique. The DNAse I-cleaved fragments were transferred onto a nitrocellulose sheet after size fractionation by gel electrophoresis. Hybridization was then carried out with radioactively labeled cloned fragments of DNA from D. melanogaster. For the 5 different chromosomal regions examined, each gives a unique pattern of higher order bands on the autoradiogram; the patterns are different for different regions. Restriction enzyme cleavage of the fragments generated indicates that the preferential DNAse I cleavage sites in chromatin are position-specific. The chromosomal regions bounded by preferential DNAse I cleavage sites are referred to as supranucleosomal or higher order domains for purposes of discussion and analysis. The micrococcal nuclease cleavage pattern of chromatin at specific loci was also examined. In 1 case, this nuclease also cleaves at position-specific sites.


Accession: 006614591



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