The chromatin structure of specific genes: I. Evidence for higher order domains of defined DNA sequence

Carl Wu*, Paul M. Bingham, Kenneth J. Livak, Robert Holmgren, Sarah C R Elgin

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

271 Scopus citations

Abstract

When the chromatin of Drosophila is examined by digestion with DNAase I or micrococcal nuclease, no general structural organization above the level of the nucleosome is revealed by the cleavage pattern. In contrast, the DNAase 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 DNAase 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 five 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 DNAase I cleavage sites in chromatin are position-specific. The chromosomal regions bounded by preferential DNAase 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 the one case studied in detail, this nuclease also cleaves at position-specific sites.

Original languageEnglish (US)
Pages (from-to)797-806
Number of pages10
JournalCell
Volume16
Issue number4
DOIs
StatePublished - Apr 1979

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

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