Loosened nucleosome linker folding in transcriptionally active chromatin of chicken embryo erythrocyte nuclei.
We have investigated the mechanism of the electrophoresis-driven chromatin aggregation which had been described by Weintraub (1984, Cell 38, 17-27) as a putative mean for propagation of genetic repression in eukaryotes. We show that the oligonucleosome aggregates are assembled de novo at the starting zone of DNP electrophoresis. A new system of native two-dimensional DNP electrophoresis has been worked out to separate the oligonucleosome aggregates ('A' particles) and the freely-migrating oligonucleosomes ('B' particles). The 'B' particle fraction which is derived from transcriptionally-active chromatin regions undergoes an extensive nuclease degradation of its DNA termini during the nuclease digestion. This fraction is partially depleted of histones H1 and H5 and is enriched in HMG nonhistone proteins. 'A' particles comprise the repressed chromatin DNA fragments which are about 60 b.p. longer than the corresponding DNA oligomers of 'B' particles. An oligonucleosome preparation containing the elongated DNA oligomers has been also isolated by means of sucrose gradient ultracentrifugation. Exonuclease III mapping reveals that the two chromatin fractions differ by an extent of terminal linker DNA trimming during the Micrococcal nuclease digestion rather than by the nucleosome repeat length. The complex character of nuclease digestion is not observed when the chromatin is digested in solution after the nuclear lysis. We argue that the protection of terminal oligonucleosome linkers is due to selective condensation of inactive chromatin in chicken erythrocyte nuclei and that the terminal DNA tails together with linker histones bound to them mediate the aggregation of repressed chromatin fragments.
Grigoryev
S A
SA
Department of Molecular Biology, Biology Faculty, Moscow State University, USSR.
Spirin
K S
KS
Krasheninnikov
I A
IA
D000818:Animals; D002642:Chick Embryo; D002843:Chromatin / Q000737:chemistry / Q000378:metabolism*; D004247:DNA / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional; D004912:Erythrocytes; D005090:Exodeoxyribonucleases / Q000378:metabolism; D008836:Micrococcal Nuclease / Q000378:metabolism; D009707:Nucleosomes / Q000378:metabolism*; D014158:Transcription, Genetic; D014461:Ultracentrifugation
D002843:Chromatin; D009707:Nucleosomes; D004247:DNA; D005090:Exodeoxyribonucleases; C020133:exodeoxyribonuclease III; D008836:Micrococcal Nuclease