DNase I and hydroxyl radical characterization of chromatin complexes.
The native chromatin complex within most eukaryotic nuclei is very difficult to study by biochemical means, so researchers have developed methods for studying smaller portions of the complex. This unit details the use of DNase I and hydroxyl radicals to characterize histone-DNA interactions within such portions of the complex. DNase I digestion can be used to determine what regions of a DNA segment are intimately associated with the core histone proteins and what regions are more like naked DNA (i.e., linker DNA within the nucleosomal repeat). The finer details of histone-DNA interactions and DNA structure within these complexes is best characterized by digestion with the hydroxyl radical. Both reagents may be used to assess the degree and homogeneity of rotational and translational positioning within isolated chromatin complexes.
Vitolo
J M
JM
University of Rochester Medical Center, Rochester, New York, USA.
Thiriet
C
C
Hayes
J J
JJ
D000818:Animals; D001345:Autoradiography; D002843:Chromatin / Q000737:chemistry* / Q000378:metabolism; D004247:DNA / Q000032:analysis* / Q000378:metabolism; D018983:DNA Footprinting; D003850:Deoxyribonuclease I / Q000032:analysis* / Q000378:metabolism; D004587:Electrophoresis, Agar Gel; D006657:Histones / Q000032:analysis*; D006801:Humans; D017665:Hydroxyl Radical / Q000737:chemistry*; D009690:Nucleic Acid Conformation; D009707:Nucleosomes / Q000737:chemistry
D002843:Chromatin; D006657:Histones; D009707:Nucleosomes; D017665:Hydroxyl Radical; D004247:DNA; D003850:Deoxyribonuclease I