Chromatin assembly during SV40 DNA replication in vitro

Stillman, B. (1986) Chromatin assembly during SV40 DNA replication in vitro. Cell, 45 (4). pp. 555-65. ISSN 0092-8674 (Print)

URL: http://www.ncbi.nlm.nih.gov/pubmed/3011272
DOI: 10.1016/0092-8674(86)90287-4

Abstract

A cytosol extract from human 293 cells supports efficient replication of SV40 origin-containing plasmid DNA in the presence of the SV40 T antigen. Addition of a nuclear extract from the same cells promotes negative supercoiling of the replicated DNA but not the bulk of the unreplicated DNA. The level of superhelicity is affected by the concentrations of T antigen and nuclear extract factors and by the time of addition of the nuclear extract. The replicated DNA in isolated DNA-protein complexes resists relaxation by purified HeLa cell topoisomerase I. Micrococcal nuclease digestion, sucrose gradient sedimentation, and electron microscopy demonstrate that the negative supercoils result from assembly of the replicating DNA into a chromatin structure. These results suggest that, during DNA replication, the core histones can be assembled on both sides of the replication fork by an active, replication-linked mechanism that does not require a template of preexisting nucleosomes.

Item Type: Paper
Additional Information:
Uncontrolled Keywords: Antigens Polyomavirus Transforming Antigens Viral, Tumor physiology Cell Line Cell Nucleus physiology Cell-Free System Chromatin ultrastructure Cytosol physiology DNA Replication DNA Topoisomerases Type I metabolism DNA, Superhelical biosynthesis DNA Viral biosynthesis Humans Oncogene Proteins Viral physiology Simian virus 40 physiology Virus Replication
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > DNA replication
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > Chromatin dynamics
CSHL Authors:
Communities: CSHL labs > Stillman lab
Highlight: Stillman, Bruce W.
Depositing User: CSHL Librarian
Date Deposited: 29 Feb 2012 19:46
Last Modified: 20 Jun 2017 20:35
Related URLs:
URI: http://repository.cshl.edu/id/eprint/25065

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