Here we describe an adapted ChIP-on-chip protocol for the analysis of DNA topoisomerase chromosomal binding in Saccharomyces cerevisiae cells. The ChIP-on-chip technique is based on the immunoprecipitation of crosslinked chromatin (ChIP, chromatin immunoprecipitation), followed by DNA amplification and hybridization to high-density oligonucleotide arrays (Chip). Comparison of the signal intensities of immunoprecipitated and control fractions provides a measurement of the protein-DNA association along entire genomes. ChIP-on-chip analysis of DNA topoisomerase binding to chromosomal DNA opens a window to the understanding of the in vivo contribution of these enzymes to the different DNA transactions taking place concomitantly within the context of the highly organized eukaryotic genome. Chromosomal binding profiles obtained from synchronized cells allow scoring the temporal and spatial restriction of these enzymes at different cell cycle stages. By using this approach, novel aspects of DNA topoisomerase function in chromosome metabolism might be unmasked.

ChIP-on-chip analysis of DNA topoisomerases / R. Bermejo, Y. Mori Katou, K. Shirahige, M. Foiani (METHODS IN MOLECULAR BIOLOGY). - In: DNA Topoisomerases: Methods And Protocols / [a cura di] DJ Clarke. - [s.l] : Humana Press, 2009. - ISBN 9781607613398. - pp. 103-118

ChIP-on-chip analysis of DNA topoisomerases

M. Foiani
Ultimo
2009

Abstract

Here we describe an adapted ChIP-on-chip protocol for the analysis of DNA topoisomerase chromosomal binding in Saccharomyces cerevisiae cells. The ChIP-on-chip technique is based on the immunoprecipitation of crosslinked chromatin (ChIP, chromatin immunoprecipitation), followed by DNA amplification and hybridization to high-density oligonucleotide arrays (Chip). Comparison of the signal intensities of immunoprecipitated and control fractions provides a measurement of the protein-DNA association along entire genomes. ChIP-on-chip analysis of DNA topoisomerase binding to chromosomal DNA opens a window to the understanding of the in vivo contribution of these enzymes to the different DNA transactions taking place concomitantly within the context of the highly organized eukaryotic genome. Chromosomal binding profiles obtained from synchronized cells allow scoring the temporal and spatial restriction of these enzymes at different cell cycle stages. By using this approach, novel aspects of DNA topoisomerase function in chromosome metabolism might be unmasked.
English
Animals; Oligonucleotide Array Sequence Analysis; Models, Genetic; Saccharomyces cerevisiae Proteins; DNA; Chromatin Immunoprecipitation; Amino Acid Sequence; Protein Binding; Nucleic Acid Conformation; DNA Topoisomerases; Saccharomyces cerevisiae; Chromosomes, Fungal
Settore BIO/11 - Biologia Molecolare
Capitolo o Saggio
Esperti anonimi
Pubblicazione scientifica
DNA Topoisomerases: Methods And Protocols
DJ Clarke
Humana Press
2009
103
118
16
9781607613398
582
Volume a diffusione internazionale
Pubmed
Aderisco
R. Bermejo, Y. Mori Katou, K. Shirahige, M. Foiani
Book Part (author)
reserved
268
ChIP-on-chip analysis of DNA topoisomerases / R. Bermejo, Y. Mori Katou, K. Shirahige, M. Foiani (METHODS IN MOLECULAR BIOLOGY). - In: DNA Topoisomerases: Methods And Protocols / [a cura di] DJ Clarke. - [s.l] : Humana Press, 2009. - ISBN 9781607613398. - pp. 103-118
info:eu-repo/semantics/bookPart
4
Prodotti della ricerca::03 - Contributo in volume
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2434/658622
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