FRET-based methods to study ATP-dependent changes in chromatin structure.
DNA packaging into chromatin imposes several levels of regulation on the central nuclear processes of DNA replication, recombination, repair and transcription. ATP-dependent chromatin-remodeling enzymes play a critical role in this regulation by altering the accessibility of nucleosomal DNA. Remodeling can result in large-scale changes in chromatin, such as the formation of heterochromatin, or smaller changes in exposure or occlusion of specific DNA regions. To understand the mechanisms of chromatin remodeling, we report a FRET-based method to follow remodeling of a single histone octamer on DNA. This technique provides a non-perturbing, solution-based approach to quantitatively track the movement of DNA with respect to the octamer in real-time. The method can easily be altered to examine other conformational changes within the nucleosome, and is applicable to study the enzymatic activity of several classes of chromatin-remodeling complexes.
Yang
Janet G
JG
Department of Biochemistry and Biophysics, University of California, San Francisco, 600 16th Street, San Francisco, CA 94158, USA.
Narlikar
Geeta J
GJ
D000255:Adenosine Triphosphate / Q000378:metabolism*; D000818:Animals; D042002:Chromatin Assembly and Disassembly / Q000502:physiology*; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D006570:Heterochromatin / Q000737:chemistry / Q000378:metabolism*; D007700:Kinetics; D009707:Nucleosomes / Q000737:chemistry / Q000378:metabolism*; D014981:Xenopus
D006570:Heterochromatin; D009707:Nucleosomes; D000255:Adenosine Triphosphate; D004247:DNA