Laser-induced crosslinking of histones to DNA in chromatin and core particles: Implications in studying histone-DNA interactions

V. Yu Stefanovsky, S. I. Dimitrov, V. R. Russanova, D. Angelov, I. G. Pashev

Research output: Contribution to journalArticle

45 Citations (Scopus)

Abstract

UV laser irradiation has been used to covalently crosslink histones to DNA in nuclei, chromatin and core particles and the presence of the different histone species in the covalently linked material was detected immunochemically. When nuclei were irradiated and then trypsinized to cleave the N- and C- terminal histone tails, no histones have been found covalently linked to DNA. This finding shows that UV laser-induced crosslinking of histories to DNA is accomplished via the non-structured domains only. This unexpected way of crosslinking operated in chromatin, H1-depleted chromatin and core particles, i.e. independently of the chromatin structure. The efficiency of crosslinking, however, showed such a dependence: whilst the yield of crosslinks was similar in total and H1-depleted chromatin, in core particles the efficiency was 3-4 times lower for H2A, H2B and H4 and 10-12 times lower for H3. The decreased crosslinking efficiency, especially dramatic in the case of 1-43, is attributed to a reduced number of binding sites, and, respectively, is considered as a direct evidence for interaction of nonstructered tails of core histones with linker DNA.

Original languageEnglish
Pages (from-to)10069-10081
Number of pages13
JournalNucleic Acids Research
Volume17
Issue number23
DOIs
Publication statusPublished - 11 Dec 1989
Externally publishedYes

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ASJC Scopus subject areas

  • Statistics, Probability and Uncertainty
  • Applied Mathematics
  • Health, Toxicology and Mutagenesis
  • Toxicology
  • Genetics(clinical)
  • Genetics

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