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PMID: 9628878 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

In vitro chromatin remodelling by chromatin accessibility complex (CHRAC) at the SV40 origin of DNA replication.

The EMBO journal ·Vol. 17 ·No. 12 ·1998-06-15 ·Pages 3428-38

Alexiadis V, Varga-Weisz PD, Bonte E, Becker PB, Gruss C

Abstract

DNA replication is initiated by binding of initiation factors to the origin of replication. Nucleosomes are known to inhibit the access of the replication machinery to origin sequences. Recently, nucleosome remodelling factors have been identified that increase the accessibility of nucleosomal DNA to transcription regulators. To test whether the initiation of DNA replication from an origin covered by nucleosomes would also benefit from the action of nucleosome remodelling factors, we reconstituted SV40 DNA into chromatin in Drosophila embryo extracts. In the presence of T-antigen and ATP, a chromatin-associated cofactor allowed efficient replication from a nucleosomal origin in vitro. In search of the energy-dependent cofactor responsible we found that purified 'chromatin accessibility complex' (CHRAC) was able to alter the nucleosomal structure at the origin allowing the binding of T-antigen and efficient initiation of replication. These experiments provide evidence for the involvement of a nucleosome remodelling machine in structural changes at the SV40 origin of DNA replication in vitro.

MeSH Terms
Animals Chromatin/genetics DNA Replication Drosophila/embryology,genetics Histones/metabolism Nucleosomes/metabolism Replication Origin/genetics Simian virus 40/genetics
Chemicals
Chromatin Histones Nucleosomes
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Alexiadis V
University of Konstanz, Division of Biology, Konstanz, Germany.
Varga-Weisz P D
Bonte E
Becker P B
Gruss C
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/17.12.3428
Published
1998-06-15
Pages
3428-38
Language
English
Region
England
NLM ID
8208664
PMCID
PMC1170679
Subset
IM
External Links
PubMed source DOI: 10.1093/emboj/17.12.3428 PMC Full Text View journal details
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