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PMID: 693319 Published · ppublish English Journal Article

Photochemical cross-linking of histones to DNA nucleosomes.

Nucleic acids research ·Vol. 5 ·No. 8 ·1978-08-00 ·Pages 2755-73

Sperling J, Sperling R

Abstract

Ultraviolet (UV)-induced cross-linking was utilized in order to identify histone-DNA interacting regions in the chromatin repeating unit. Fractionated mononucleosomes which contained 185 base pairs of DNA and a full complement of the histones, including histone H1, were irradiated with light of lambda greater than 290nm in the presence of a photosensitizer. Equimolar amounts of histones H2A and H2B were found, by two independent labeling experiments, to be cross-linked to the DNA. Based on previous finding that the UV irradiation specifically cross-links residues which are in close proximity, irrespective of the nature of the amino acid side chain or the nucleotide involved, our results indicate that the four core histones are not positioned equivalently with respect to the DNA. This arrangement allows histones H2A and H2B to preferentially cross-link to the DNA. A water soluble covalent complex of DNA and histones was isolated. This complex was partially resistant to mild nuclease digestion, it exhibited a CD spectrum similar to that of chromatin, and was found to contain histone H1. These results are compatible with a model which suggests that histone H1, though anchored to the linker, is bound to the DNA at additional sites. By doing so it spans the whole length of the nucleosome and clamps together the DNA fold around the histone core.

MeSH Terms
Animals Binding Sites Chromatin/radiation effects,ultrastructure DNA/radiation effects Histones/radiation effects Liver/ultrastructure Photochemistry Protein Binding Rats Ultraviolet Rays
Chemicals
Chromatin Histones DNA
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Sperling J
Sperling R
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51 references, click to expand
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/5.8.2755
Published
1978-08-00
Pages
2755-73
Language
English
Region
England
NLM ID
0411011
PMCID
PMC342205
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/5.8.2755 PMC Full Text View journal details
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