Characteristics of chromatin release during digestion of nuclei with micrococcal nuclease: preferential solubilization of nascent RNA at low enzyme concentration.
1. Extensive digestion of nuclei with micrococcal nuclease (MNase), commonly used in the analysis of chromatin structure, results in the production of mono- and dinucleosomal chromatin fragments. 2. Digestion of nuclei from a range of cell types with low enzyme concentrations solubilized high molecular weight polynucleosomal fragments, some greater than 22 kb long. 3. Such digestion conditions also resulted in extensive solubilization of nascent RNA which contributed considerably to the nucleic acid content of the soluble fraction. 4. We conclude that the contribution of RNA to total nucleic acid content of the soluble fraction should be taken into consideration when nuclei are digested with low concentrations of MNase.
Telford
D J
DJ
Children's Leukaemia and Cancer Research Unit, Prince of Wales Children's Hospital, Randwick, NSW, Australia.
Stewart
B W
BW
D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism*; D002843:Chromatin / Q000378:metabolism*; D004247:DNA / Q000378:metabolism; D006801:Humans; D015473:Leukemia, Promyelocytic, Acute; D008099:Liver / Q000648:ultrastructure; D008836:Micrococcal Nuclease / Q000378:metabolism*; D008970:Molecular Weight; D012313:RNA / Q000378:metabolism*; D051381:Rats; D012175:Retinoblastoma; D012995:Solubility; D014407:Tumor Cells, Cultured
D002843:Chromatin; D012313:RNA; D004247:DNA; D008836:Micrococcal Nuclease