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

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 si...

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Bibliographic Details
Published in:Nucleic acids research 1988-05, Vol.16 (10), p.4525-4538
Main Authors: ANGELOV, D, STEFANOVSKY, V. YU, DIMITROV, S. I, RUSSANOVA, V. R, KESKINOVA, E, PASHEV, I. G
Format: Article
Language:English
Subjects:
Animals
Biological and medical sciences
Carbon Radioisotopes
Carcinoma, Ehrlich Tumor - metabolism
Cell Nucleus - metabolism
Cell Nucleus - radiation effects
Chickens
Chromatin. Chromosome
Cross-Linking Reagents
DNA - blood
DNA - radiation effects
Erythrocytes - metabolism
Female
Fundamental and applied biological sciences. Psychology
Histones - metabolism
Histones - radiation effects
Kinetics
Mice
Molecular and cellular biology
Molecular genetics
Nuclear Proteins - metabolism
Nuclear Proteins - radiation effects
Time Factors
Ultraviolet Rays
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Summary: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.
ISSN:0305-1048
1362-4962
DOI:10.1093/nar/16.10.4525