Picosecond laser cross: linking histones to DNA in chromatin: implication in studying histone/DNA interactions

Dimitar A. Angelov; S. Dimitrov; E. Keskinova; I. Pashev; V. Russanova; Yu. Stefanovsky

doi:10.1117/12.57287

1 May 1991 Picosecond laser cross, linking histones to DNA in chromatin: implication in studying histone-DNA interactions

Dimitar A. Angelov, S. Dimitrov, E. Keskinova, I. Pashev, V. Russanova, Yu. Stefanovsky

Author Affiliations +

Proceedings Volume 1403, Laser Applications in Life Sciences; (1991) https://doi.org/10.1117/12.57287
Event: Laser Applications in Life Sciences, 1990, Moscow, Russian Federation

Abstract

A picosecond UV laser radiation was used to cross-link proteins to DNA in nuclei, whole cells and different chromating preparations. All histones as well as high-mobility group 1 proten were identified immunochemically in the covalently linked protein-DNA complexes. Irradiation of the nuclieohistone resulted in cross-linking 20% of bound histones to DNA as a result of two-quantum photoreaction with a maximum quantum yield 3.10 -4 for double stranded DNA. When nuclei, total bromatin Hi-depleted chromatin and core particles were irradiated and then trypsinized or treated with clostripain to cleave respectively the N-, C- and N- terminal histone tails, no histones have been found covalently linked to DNA. However whilst the yield of cross-links was similar in total and H1-depleted chromatin in core particples the efficiencey was 3-4 times lower for H2A, H2B and H4 10-12 times lower for H3. This finding we consider as a direct evidence for interaction of non structured N- tails of core histones with linker DNA. Cross-linking in core particles depends on the ionic strength. All histones were identified in the complex formed up to 0.4 N NaCl, no cross-linking was observed when irradiation was carried out at salt concentration higher than 0.4 M. The cross-linking ability was preserved both upon physiological acetylation of histones knows to be restriced to the N-terminal tails and with chemically acetylated chromatin. This finding is direct evidence that postsynthetic histone acetylation does not release the N-terminal tails from interaction with DNA.

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Dimitar A. Angelov, S. Dimitrov, E. Keskinova, I. Pashev, V. Russanova, and Yu. Stefanovsky "Picosecond laser cross, linking histones to DNA in chromatin: implication in studying histone-DNA interactions", Proc. SPIE 1403, Laser Applications in Life Sciences, (1 May 1991); https://doi.org/10.1117/12.57287

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KEYWORDS

Particles

Proteins

Picosecond phenomena

Laser applications

Ultraviolet radiation

Photons

Chlorine

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