Analysis of histone deposition on specific DNA elements in living mammalian cells.
DNA replication and repair involve the deposition of newly synthesized histones. After their deposition on a specific locus, histones are extensively modified on their protruding N-terminal tails to restore the histone code corresponding to this locus. These processes require a complex machinery of histone chaperones and histone-modifying enzymes. The precise kinetics of these events and the identity of the factors involved are still not clear. Here we present a technique based on transient expression of tagged histones followed by chromatin immunoprecipitation that allows the detection of newly synthesized histone deposition on specific DNA elements in mammalian cells at high resolution. This strategy could have important applications in chromatin dynamic studies.
Daury
Laetitia
L
Laboratoire de Biologie Moléculaire Eucaryote, NRS UMR 5099, Ligue Nationale Contre le Cancer, Toulouse, France.
Trouche
Didier
D
D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D002477:Cells / Q000378:metabolism*; D002843:Chromatin / Q000378:metabolism; D004247:DNA / Q000235:genetics / Q000378:metabolism*; D020224:Expressed Sequence Tags; D005347:Fibroblasts / Q000378:metabolism*; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D006388:Hemagglutinins / Q000378:metabolism; D006657:Histones / Q000096:biosynthesis* / Q000737:chemistry; D008164:Luminescent Proteins; D008322:Mammals; D051379:Mice; D041681:NIH 3T3 Cells; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011233:Precipitin Tests; D011401:Promoter Regions, Genetic; D034622:RNA Interference; D013762:Tetrahydrofolate Dehydrogenase / Q000235:genetics; D014162:Transfection
D002843:Chromatin; D006388:Hemagglutinins; D006657:Histones; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D004247:DNA; D013762:Tetrahydrofolate Dehydrogenase