Protein—DNA crosslinking in reconstituted nucleohistone, nuclei and whole cells by picosecond UV laser irradiation (original) (raw)
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Institute of Solid State Physics, Bulgarian Academy of Sciences
1113 Sofia, Bulgaria
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1
Institute of Solid State Physics, Bulgarian Academy of Sciences
1113 Sofia, Bulgaria
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* To whom correspondence should be addressed
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Received:
11 December 1987
Revision received:
14 April 1988
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D. Angelov, V.Yu. Stefanovsky, S.I. Dimitrov, V.R. Russanova, E. Keskinova, I.G. Pashev, Protein—DNA crosslinking in reconstituted nucleohistone, nuclei and whole cells by picosecond UV laser irradiation, Nucleic Acids Research, Volume 16, Issue 10, 25 May 1988, Pages 4525–4538, https://doi.org/10.1093/nar/16.10.4525
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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.
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