Histone deposition and chromatin assembly by RSF.
It is becoming clear that the structure of cellular chromatin is dynamic and capable of undergoing rapid changes to respond to the metabolic requirements of the cell. These changes have a direct impact on gene expression and, therefore, the chromatin context must be considered when biochemical reactions that involve DNA are studied. Over the past several decades, a number of methods for assembling chromatin in vitro have been described. Some of them use chemical compounds to deposit histone octamers onto the DNA. Others take advantage of cellular protein complexes that have the ability to assemble chromatin. Some of these complexes have been identified and purified. This article focuses on one of these factors, RSF (remodeling and spacing factor), which was identified in our laboratory. We describe how the chromatin assembly reaction is performed and how it can be monitored to evaluate its efficiency.
Loyola
Alejandra
A
Department of Biochemistry, Division of Nucleic Acids Enzymology, Howard Hughes Medical Institute, University of Medicine and Dentistry of New Jersey, Robert Wood Johnson Medical School, Piscataway, NJ 08854, USA.
Reinberg
Danny
D
D002458:Cell Fractionation / Q000379:methods; D002467:Cell Nucleus / Q000648:ultrastructure; D002843:Chromatin / Q000648:ultrastructure*; D002848:Chromatography, DEAE-Cellulose / Q000379:methods; D004247:DNA / Q000235:genetics; D019281:Dimerization; D006367:HeLa Cells; D006657:Histones / Q000737:chemistry / Q000302:isolation & purification*; D006801:Humans; D007202:Indicators and Reagents; D008836:Micrococcal Nuclease; D008854:Microscopy, Electron; D021122:Protein Subunits / Q000302:isolation & purification
D002843:Chromatin; D006657:Histones; D007202:Indicators and Reagents; D021122:Protein Subunits; D004247:DNA; D008836:Micrococcal Nuclease