Transcription of dinucleosomal templates.
Nucleosomes and the chromatin structures they assemble provide the architectural foundation for the transcription process. We describe the use of reconstituted chromatin templates assembled using purified histones and HMGs to investigate the significance of chromatin structure for transcription. Our structural assays include sucrose gradient fractionation, nucleoprotein gel analysis, micrococcal nuclease digestion, DNase I and hydroxyl radical cleavage mapping of nucleosome position, and determination of nucleosome mobility. We determine the consequences of incorporation of linker histones or HMG1 on the properties of the nucleosome and the transcription process. Our results indicate that linker histones and HMG1 can direct the positioning of core histone-DNA interactions, restrict nucleosome mobility, and repress transcription.
Wolffe
A P
AP
Laboratory of Molecular Embryology, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20892-5430, USA.
Ura
K
K
D000818:Animals; D002499:Centrifugation, Density Gradient; D002843:Chromatin / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D018983:DNA Footprinting; D004268:DNA-Binding Proteins / Q000378:metabolism; D003850:Deoxyribonuclease I / Q000378:metabolism; D004587:Electrophoresis, Agar Gel; D004591:Electrophoresis, Polyacrylamide Gel; D006609:High Mobility Group Proteins / Q000378:metabolism; D006657:Histones / Q000302:isolation & purification / Q000378:metabolism; D017665:Hydroxyl Radical / Q000378:metabolism; D008836:Micrococcal Nuclease / Q000378:metabolism; D009707:Nucleosomes / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D015183:Restriction Mapping; D013698:Templates, Genetic; D014158:Transcription, Genetic; D014982:Xenopus laevis
D002843:Chromatin; D004268:DNA-Binding Proteins; D006609:High Mobility Group Proteins; D006657:Histones; D009707:Nucleosomes; D017665:Hydroxyl Radical; D004247:DNA; D003850:Deoxyribonuclease I; D008836:Micrococcal Nuclease