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PMID: 6283130 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Mapping the in vivo arrangement of nucleosomes on simian virus 40 chromatin by the photoaddition of radioactive hydroxymethyltrimethylpsoralen.

Journal of virology ·Vol. 41 ·No. 1 ·1982-01-00 ·Pages 78-87

Robinson GW, Hallick LM

Abstract

Intracellular simian virus 40 (SV40) chromatin was photoreacted with a 3H-labeled psoralen derivative, hydroxymethyltrimethylpsoralen (HMT), at 48 h postinfection. Psoralen compounds have been shown to readily penetrate intact cells and, in the presence of long-wavelength UV light, form covalent adducts to DNA, preferentially at regions unprotected by nucleosomes. The average distribution pattern of [3H]HMT on the SV40 genome was determined by specific activity measurements of the DNA fragments generated by HindIII plus HpaII or by AtuI restriction enzyme digestion. At levels of 1 to 10 [3H]HMT photoadducts per SV40 molecule, the radiolabel was found to be distributed nonrandomly. Comparison of the labeling pattern in vivo with that of purified SV40 DNA labeled in vitro revealed one major difference. A region of approximately 400 base pairs, located between 0.65 and 0.73 on the physical map, was preferentially labeled under in vivo conditions. This finding strongly suggests that the highly accessible region near the origin of replication, previously observed on isolated SV40 "minichromosomes," exists on SV40 chromatin in vivo during a lytic infection.

MeSH Terms
Binding Sites Chromatin/ultrastructure Chromosome Mapping/methods DNA Replication DNA Restriction Enzymes DNA, Viral/genetics Nucleosomes/ultrastructure Photochemistry Simian virus 40/genetics,ultrastructure Trioxsalen/analogs & derivatives
Chemicals
Chromatin DNA, Viral Nucleosomes hydroxymethyltrioxsalen DNA Restriction Enzymes Trioxsalen
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Robinson G W
Hallick L M
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.41.1.78-87.1982
Published
1982-01-00
Pages
78-87
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC256727
Subset
IM
Grants
NCI NIH HHS · CA 24799 · United States
NIGMS NIH HHS · GM 07336 · United States
External Links
PubMed source DOI: 10.1128/JVI.41.1.78-87.1982 PMC Full Text View journal details
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