Reconstitution of nucleosomes containing dinitrophenylated histones.
Histone octamers of purified monomer nucleosomes were labelled with [3H]dinitrofluorobenzene. Authentic 11 S nucleosomes were reconstituted in vitro from a mixture of [3H]dinitrophenylated histones and excess unlabelled monomer nucleosomes. The reconstituted nucleosomes were found to contain [3H]dinitrophenylated histones H2a and H2b but not [3H]dinitrophenylated histones H3 and H4. Approx. 83% of [3H]dinitrophenylated nucleosomes were immunoprecipitable with anti-dinitrophenyl immunoglobulin and Staphylococcus aureus. These results demonstrate that histones H2a and H2b contain dinitrofluorobenzene-reactive groups that can be modified without destroying their ability to participate in nucleosome formation in vitro.
D002499:Centrifugation, Density Gradient; D011232:Chemical Precipitation; D004139:Dinitrofluorobenzene / Q000276:immunology; D004591:Electrophoresis, Polyacrylamide Gel; D006657:Histones / Q000276:immunology; D009578:Nitrobenzenes / Q000276:immunology; D009707:Nucleosomes / Q000276:immunology
D006657:Histones; D009578:Nitrobenzenes; D009707:Nucleosomes; D004139:Dinitrofluorobenzene