RT Journal Article
SR Electronic
T1 UV Laser Cross-linking: A Real-Time Assay to Study Dynamic Protein/DNA Interactions During Chromatin Remodeling
JF Science's STKE
JO Sci. STKE
FD American Association for the Advancement of Science
SP pl13
OP pl13
DO 10.1126/stke.2562004pl13
VO 2004
IS 256
A1 Nagaich, Akhilesh K.
A1 Hager, Gordon L.
YR 2004
UL http://stke.sciencemag.org/content/2004/256/pl13.abstract
AB We describe the use of laser ultraviolet (UV) cross-linking to study the interaction of transcription factors with in vitro assembled chromatinized DNA templates in real time. Because the laser source delivers a high density of photons in a single ns pulse, the cross-linking reaction is completed in less than 1 microseconds, allowing the investigator to freeze rapid dynamic changes in protein-DNA interactions. Using this approach, we have sampled the dynamic equilibrium of the glucocorticoid receptor (GR) and the chromatin remodeling complex (SWI/SNF) during adenosine triphosphate (ATP)–dependent chromatin remodeling on a chromatinized mouse mammary tumor virus promoter in vitro. UV laser cross-linking shows that the GR and SWI/SNF complex undergoes a periodic binding and displacement event during the process of chromatin remodeling. The assay provides unique information regarding the equilibrium of protein-DNA interactions in real time and can be easily adapted to study the dynamic events in the assembly and disassembly of other multiprotein complexes on chromatin or DNA templates.