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. 1988 May 25;16(10):4525–4538. doi: 10.1093/nar/16.10.4525

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

D Angelov 1, Stefanovsky VYu 1, S I Dimitrov 1, V R Russanova 1, E Keskinova 1, I G Pashev 1
PMCID: PMC336646  PMID: 3380686

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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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Arbieva Z. Kh, Kalmykov P. V., Dobrov E. N., Esenaliev R. O., Morev P. G. Obrazovanie sshivok s belkom i razryvov polinukleotidnoi tsepi v RNK VTM in situ pod deistviem vysokointensivnogo pikosekundnogo lazernogo UF-izlucheniia. Dokl Akad Nauk SSSR. 1987 Jan-Feb;292(1):227–230. [PubMed] [Google Scholar]
  2. Budowsky E. I., Axentyeva M. S., Abdurashidova G. G., Simukova N. A., Rubin L. B. Induction of polynucleotide-protein cross-linkages by ultraviolet irradiation. Peculiarities of the high-intensity laser pulse irradiation. Eur J Biochem. 1986 Aug 15;159(1):95–101. doi: 10.1111/j.1432-1033.1986.tb09837.x. [DOI] [PubMed] [Google Scholar]
  3. Dizdaroglu M., Simic M. G. Radiation-induced crosslinks between thymine and phenylalanine. Int J Radiat Biol Relat Stud Phys Chem Med. 1985 Jan;47(1):63–69. doi: 10.1080/09553008514550091. [DOI] [PubMed] [Google Scholar]
  4. Harrison C. A., Turner D. H., Hinkle D. C. Laser crosslinking of E. coli RNA polymerase and T7 DNA. Nucleic Acids Res. 1982 Apr 10;10(7):2399–2414. doi: 10.1093/nar/10.7.2399. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Hillel Z., Wu C. W. Photochemical cross-linking studies on the interaction of Escherichia coli RNA polymerase with T7 DNA. Biochemistry. 1978 Jul 25;17(15):2954–2961. doi: 10.1021/bi00608a003. [DOI] [PubMed] [Google Scholar]
  6. Hockensmith J. W., Kubasek W. L., Vorachek W. R., von Hippel P. H. Laser cross-linking of nucleic acids to proteins. Methodology and first applications to the phage T4 DNA replication system. J Biol Chem. 1986 Mar 15;261(8):3512–3518. [PubMed] [Google Scholar]
  7. Jellinek T., Johns R. B. The mechanism of photochemical addition of cysteine to uracil and formation of dihydrouracil. Photochem Photobiol. 1970 May;11(5):349–359. doi: 10.1111/j.1751-1097.1970.tb06007.x. [DOI] [PubMed] [Google Scholar]
  8. Kunkel G. R., Martinson H. G. Histone-DNA interactions within chromatin. Isolation of histones from DNA-histone adducts induced in nuclei by UV light. Nucleic Acids Res. 1978 Nov;5(11):4263–4272. doi: 10.1093/nar/5.11.4263. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  10. Lica L., Ray D. S. Replication of bacteriophage M13. XII. In vivo cross-linking of a phage-specific DNA binding protein to the single-stranded DNA of bacteriophage M13 by ultraviolet irradiation. J Mol Biol. 1977 Sep;115(1):45–59. doi: 10.1016/0022-2836(77)90245-5. [DOI] [PubMed] [Google Scholar]
  11. Markovitz A. Ultraviolet light-induced stable complexes of DNA and DNA polymerase. Biochim Biophys Acta. 1972 Nov 9;281(4):522–534. doi: 10.1016/0005-2787(72)90153-0. [DOI] [PubMed] [Google Scholar]
  12. Messing J. New M13 vectors for cloning. Methods Enzymol. 1983;101:20–78. doi: 10.1016/0076-6879(83)01005-8. [DOI] [PubMed] [Google Scholar]
  13. Nikogosyan D. N., Angelov D. A., Oraevsky A. A. Determination of parameters of excited states of DNA and RNA bases by laser UV photolysis. Photochem Photobiol. 1982 May;35(5):627–635. doi: 10.1111/j.1751-1097.1982.tb02621.x. [DOI] [PubMed] [Google Scholar]
  14. Pashev I. G., Dimitrov S. I., Ivanov I. G., Markov G. G. Histone acetylation in chromatin containing mouse satellite DNA. Eur J Biochem. 1983 Jun 15;133(2):379–382. doi: 10.1111/j.1432-1033.1983.tb07473.x. [DOI] [PubMed] [Google Scholar]
  15. Pashev I. G., Nencheva M. M., Markov G. G. Denaturation of mouse satellite DNA upon melting of chromatin in solution. Mol Biol Rep. 1978 Oct 16;4(3):143–147. doi: 10.1007/BF00777514. [DOI] [PubMed] [Google Scholar]
  16. Russanova V. R., Dimitrov S. I., Makarov V. L., Pashev I. G. Accessibility of the globular domain of histones H1 and H5 to antibodies upon folding of chromatin. Eur J Biochem. 1987 Sep 1;167(2):321–326. doi: 10.1111/j.1432-1033.1987.tb13339.x. [DOI] [PubMed] [Google Scholar]
  17. Smith K. C. A mixed photoproduct of thymine and cysteine: 5-S-cysteine, 6-hydrothymine. Biochem Biophys Res Commun. 1970;39(6):1011–1016. doi: 10.1016/0006-291x(70)90658-3. [DOI] [PubMed] [Google Scholar]
  18. Strniste G. F., Smith D. A. Induction of stable linkage between the deoxyribonucleic acid dependent ribonucleic acid polymerase and d(A-T)n-d(A-T)n by ultraviolet light. Biochemistry. 1974 Jan 29;13(3):485–493. doi: 10.1021/bi00700a014. [DOI] [PubMed] [Google Scholar]

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