Non-histone proteins of soluble nucleoproteins released from mouse myeloma nuclei by mild micrococcal nuclease digestion.
Mono- and dinucleosomes preferentially cleaved from mouse myeloma chromatin by very mild micrococcal nuclease digestion at 0 degree C are soluble and are released from nuclei under near-physiological conditions in which normal nucleosomes containing Hl are insoluble. These nucleosomes are highly enriched in RNA, high-mobility-group proteins and a unique subset of other non-histone proteins. They are nearly devoid of histone Hl and contain DNA significantly less methylated than whole myeloma DNA, indicating that they comprise a subset of genomic sequences. Previously we have shown that this fraction is enriched in transcribed DNA sequences. Non-histone proteins that co-sedimented with readily solubilized nucleosomes included many of the most basic, low-to-moderate molecular weight chromosomal proteins. Many of these proteins were also preferentially acetylated in vivo. The residual, pelleted chromatin was highly enriched in high molecular weight proteins (greater than 60 000), and very depleted in medium molecular weight proteins. Readily solubilized nucleoproteins sedimenting like mononucleosomes were partly resolved by electrophoresis, under non-denaturing conditions, into several subfractions differing significantly in non-histone protein contents. Methods described here should be useful for identifying and isolating non-histone proteins bound to nucleosomes and other chromatin regions that are structurally and functionally unique.
D000085:Acetates / Q000378:metabolism; D000818:Animals; D002250:Carbon Radioisotopes; D002460:Cell Line; D002467:Cell Nucleus / Q000648:ultrastructure*; D002868:Chromosomal Proteins, Non-Histone / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel; D051379:Mice; D008807:Mice, Inbred BALB C; D008836:Micrococcal Nuclease / Q000378:metabolism*; D008970:Molecular Weight; D009698:Nucleoproteins / Q000302:isolation & purification*; D010954:Plasmacytoma / Q000032:analysis*
D000085:Acetates; D002250:Carbon Radioisotopes; D002868:Chromosomal Proteins, Non-Histone; D009698:Nucleoproteins; D008836:Micrococcal Nuclease