The fate of parental nucleosomes during SV40 DNA replication.
The fate of parental nucleosomes during the replication of chromatin templates was studied using a modification of the cell-free SV40 DNA replication system. Plasmid DNA molecules containing the SV40 origin were assembled into chromatin with purified core histones and fractionated assembly factors derived from HeLa cells. When these templates were replicated in vitro, the resulting progeny retained a nucleosomal organization. To determine whether the nucleosomes associated with the progeny molecules resulted from displacement of parental histones during replication followed by reassembly, the replication reactions were performed in the presence of control templates. It was observed that the progeny genomes resulting from the replication of chromatin templates retained a nucleosomal structure, whereas the progeny of the control DNA molecules were not assembled into chromatin. Additional experiments, involving direct addition of histones to the replication reaction mixtures, confirmed that the control templates were not sequestered in some form which made them unavailable for nucleosome assembly. Thus, our data demonstrate that parental nucleosomes remain associated with the replicating molecules and are transferred to the progeny molecules without displacement into solution. We propose a simple model in which nucleosomes ahead of the fork are transferred intact to the newly synthesized daughter duplexes.
Randall
S K
SK
Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.
Kelly
T J
TJ
D002474:Cell-Free System; D002843:Chromatin / Q000502:physiology; D004261:DNA Replication; D004279:DNA, Viral / Q000235:genetics*; D016679:Genome, Viral; D006367:HeLa Cells; D006657:Histones / Q000302:isolation & purification / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D008836:Micrococcal Nuclease; D009707:Nucleosomes / Q000502:physiology*; D015183:Restriction Mapping; D013539:Simian virus 40 / Q000235:genetics*; D013698:Templates, Genetic
D002843:Chromatin; D004279:DNA, Viral; D006657:Histones; D009707:Nucleosomes; D008836:Micrococcal Nuclease