Transcriptionally active chromatin is sensitive to Neurospora crassa and S1 nucleases.
We have examined the distribution of Neurospora crassa and S1 nuclease cleavage products in the chromatin of the 87A7 heat shock locus of Drosophila melanogaster. Both of these nucleases generate single and double-strand breaks in chromatin at specific sites in the 87A7 locus. Before heat induction, we find that the 5' ends of the two 87A7 hsp 70 genes contain N. crassa and S1 nuclease hypersensitive sites, while there are only a few cleavage products from elsewhere in the locus. With N. crassa nuclease, we observe one major 5' fragment, and this is derived from cleavage in a DNA segment mapping about 90 to 115 base-pairs from the beginning of the transcription unit. With S1 nuclease, we find two 5' cleavage products. The first maps about 120 to 130 base-pairs from the beginning of the gene. Interestingly, this site is also sensitive to S1 nuclease in supercoiled but not linear naked DNA. The other fragment maps very close to the transcription start site (approximately 0 to -15 base-pairs). After heat induction, there is a transition in the chromatin architecture of 87A7. First, there is a marked reduction in the yield of the prominent 5' N. crassa and S1 nuclease fragments. Second, the entire hsp 70 gene, as well as the spacer DNA just downstream from the 3' end of the gene, becomes highly sensitive to both of these nucleases.
D000818:Animals; D002843:Chromatin / Q000235:genetics; D002874:Chromosome Mapping; D004247:DNA; D004262:DNA Restriction Enzymes; D004277:DNA, Single-Stranded; D004278:DNA, Superhelical; D004331:Drosophila melanogaster; D004587:Electrophoresis, Agar Gel; D004720:Endonucleases; D005796:Genes; D006360:Heat-Shock Proteins / Q000235:genetics; D006358:Hot Temperature; D015719:Single-Strand Specific DNA and RNA Endonucleases; D014158:Transcription, Genetic
D002843:Chromatin; D004277:DNA, Single-Stranded; D004278:DNA, Superhelical; D006360:Heat-Shock Proteins; D004247:DNA; D004720:Endonucleases; D004262:DNA Restriction Enzymes; D015719:Single-Strand Specific DNA and RNA Endonucleases