Protein - DNA crosslinking in reconstituted nucleohistone, nuclei and whole cells by picosecond UV laser irradiation

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

Research output: Contribution to journalArticle

46 Citations (Scopus)

Abstract

A picosecond UV laser was used to cross-link proteins to DNA in nuclei, whole cells and reconstituted nucleohistone. Irradiation of the nucleohistone resulted in crosslinking 15-20% of bound histones to DNA in a very short time (one or several picosecond pulses), the efficiency of crosslinking to single stranded DNA being higher than to double stranded DNA. All histones as well as high mobility group 1 proteins were identified in the covalently linked protein-DNA complexes upon irradiation of isolated nuclei and whole cells. A method is suggested for isolation of crosslinked material from cells and nuclei in amounts sufficient for further analysis. Experiments with reconstituted nucleohistones showed that upon irradiation at a constant dose the efficiency of crosslinking depended on the intensity of the light, thus suggesting a two-quantum process is involved in the reaction.

Original languageEnglish
Pages (from-to)4525-4538
Number of pages14
JournalNucleic Acids Research
Volume16
Issue number10
DOIs
Publication statusPublished - 25 May 1988
Externally publishedYes

Fingerprint

UV Laser
Laser beam effects
Cell Nucleus
Irradiation
Crosslinking
Nucleus
Lasers
DNA
Proteins
Protein
Cell
Histones
High Mobility Group Proteins
Isolation
Single-Stranded DNA
Dose
Sufficient
Dosimetry
Light
Experiment

ASJC Scopus subject areas

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

Cite this

Protein - DNA crosslinking in reconstituted nucleohistone, nuclei and whole cells by picosecond UV laser irradiation. / Angelov, D.; Stefanovsky, V. Yu; Dimitrov, S. I.; Russanova, V. R.; Keskinova, E.; Pashev, I. G.

In: Nucleic Acids Research, Vol. 16, No. 10, 25.05.1988, p. 4525-4538.

Research output: Contribution to journalArticle

Angelov, D. ; Stefanovsky, V. Yu ; Dimitrov, S. I. ; Russanova, V. R. ; Keskinova, E. ; Pashev, I. G. / Protein - DNA crosslinking in reconstituted nucleohistone, nuclei and whole cells by picosecond UV laser irradiation. In: Nucleic Acids Research. 1988 ; Vol. 16, No. 10. pp. 4525-4538.
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