UV-laser crosslinking of proteins to DNA - PubMed (original) (raw)
UV-laser crosslinking of proteins to DNA
T Moss et al. Methods. 1997 Feb.
Abstract
Photochemical crosslinking is now a powerful method for studying protein-nucleic acid interactions. UV light is a zero-length crosslinking agent that predominantly or exclusively crosslinks proteins to nucleic acids at their contact points. It can therefore provide strong evidence for close protein-nucleic acid interactions. However, to achieve an acceptable degree of crosslinking with conventional UV light sources, exposure times ranging from minutes to several hours are necessary. Such prolonged irradiation allows for the artifactual redistribution of proteins and precludes kinetic studies. The use of UV lasers overcomes these difficulties since the number of photons required for the crosslinking may be delivered in time intervals on the order of nano- or even picoseconds. We described detailed procedures for UV laser-induced protein-DNA crosslinking both in vivo and in vitro. Technical aspects, including the choice of UV laser for irradiation, the isolation of covalently crosslinked protein-DNA complexes, immunochemical techniques for both the identification and isolation of specific protein-DNA complexes and the identification of the crosslinked DNA sequences, are reviewed in detail. The application of UV laser crosslinking in kinetic studies is illustrated by the example of the TATA-binding protein (TBP) interaction with the adenovirus E4 promoter.
Similar articles
- Detecting DNA-binding of proteins in vivo by UV-crosslinking and immunoprecipitation.
Zhang L, Zhang K, Prändl R, Schöffl F. Zhang L, et al. Biochem Biophys Res Commun. 2004 Sep 24;322(3):705-11. doi: 10.1016/j.bbrc.2004.07.202. Biochem Biophys Res Commun. 2004. PMID: 15336521 Review. - Detection and isolation of DNA-binding proteins using single-pulse ultraviolet laser crosslinking.
Ho DT, Sauvé DM, Roberge M. Ho DT, et al. Anal Biochem. 1994 May 1;218(2):248-54. doi: 10.1006/abio.1994.1174. Anal Biochem. 1994. PMID: 8074276 - Comparison of femtosecond laser and continuous wave UV sources for protein-nucleic acid crosslinking.
Fecko CJ, Munson KM, Saunders A, Sun G, Begley TP, Lis JT, Webb WW. Fecko CJ, et al. Photochem Photobiol. 2007 Nov-Dec;83(6):1394-404. doi: 10.1111/j.1751-1097.2007.00179.x. Photochem Photobiol. 2007. PMID: 18028214 - Signals for TBP/TATA box recognition.
Bareket-Samish A, Cohen I, Haran TE. Bareket-Samish A, et al. J Mol Biol. 2000 Jun 16;299(4):965-77. doi: 10.1006/jmbi.2000.3797. J Mol Biol. 2000. PMID: 10843851 - UV laser footprinting and protein-DNA crosslinking. Application to chromatin.
Angelov D, Khochbin S, Dimitrov S. Angelov D, et al. Methods Mol Biol. 1999;119:481-95. doi: 10.1385/1-59259-681-9:481. Methods Mol Biol. 1999. PMID: 10804534 Review. No abstract available.
Cited by
- Synthesis and polymerase bypass studies of DNA-peptide and DNA-protein conjugates.
Pujari SS, Tretyakova N. Pujari SS, et al. Methods Enzymol. 2021;661:363-405. doi: 10.1016/bs.mie.2021.09.005. Epub 2021 Oct 26. Methods Enzymol. 2021. PMID: 34776221 Free PMC article. - Atomic-resolution mapping of transcription factor-DNA interactions by femtosecond laser crosslinking and mass spectrometry.
Reim A, Ackermann R, Font-Mateu J, Kammel R, Beato M, Nolte S, Mann M, Russmann C, Wierer M. Reim A, et al. Nat Commun. 2020 Jun 15;11(1):3019. doi: 10.1038/s41467-020-16837-x. Nat Commun. 2020. PMID: 32541649 Free PMC article. - Mass Spectrometry-Based Tools to Characterize DNA-Protein Cross-Linking by Bis-Electrophiles.
Groehler A 4th, Degner A, Tretyakova NY. Groehler A 4th, et al. Basic Clin Pharmacol Toxicol. 2017 Sep;121 Suppl 3(Suppl 3):63-77. doi: 10.1111/bcpt.12751. Epub 2017 Mar 14. Basic Clin Pharmacol Toxicol. 2017. PMID: 28032943 Free PMC article. Review. - Efficiency and pattern of UV pulse laser-induced RNA-RNA cross-linking in the ribosome.
Shapkina T, Lappi S, Franzen S, Wollenzien P. Shapkina T, et al. Nucleic Acids Res. 2004 Mar 3;32(4):1518-26. doi: 10.1093/nar/gkh320. Print 2004. Nucleic Acids Res. 2004. PMID: 14999094 Free PMC article. - Localization of proteins bound to a replication origin of human DNA along the cell cycle.
Abdurashidova G, Danailov MB, Ochem A, Triolo G, Djeliova V, Radulescu S, Vindigni A, Riva S, Falaschi A. Abdurashidova G, et al. EMBO J. 2003 Aug 15;22(16):4294-303. doi: 10.1093/emboj/cdg404. EMBO J. 2003. PMID: 12912926 Free PMC article.