Single-pair Fluorescence Resonance Energy Transfer study of mononucleosomes dynamics
| Autor: | Jens Michaelis, Josef H. Exler, Barbara Treutlein, Gernot Längst |
|---|---|
| Rok vydání: | 2009 |
| Předmět: |
0303 health sciences
biology Base pair viruses Biophysics DNA replication 7. Clean energy Linker DNA Chromatin 03 medical and health sciences Crystallography chemistry.chemical_compound 0302 clinical medicine Histone chemistry biology.protein Nucleosome Histone octamer 030217 neurology & neurosurgery DNA 030304 developmental biology |
| Zdroj: | Biophysical Journal. 96:292a |
| ISSN: | 0006-3495 |
| DOI: | 10.1016/j.bpj.2008.12.1447 |
| Popis: | Nucleosomes, the primary repeating unit of chromatin, package DNA by wrapping 147 base pairs tightly around an octamer of histone proteins in approximately 1.7 helical turns. In order to allow central nuclear processes to happen, such as DNA replication, recombination, repair and transcription, regulated exposure of the template DNA is required. Therefore, the nucleosome has to undergo certain conformational changes. One possible strategy for such a regulated exposure is the translocation or ‘sliding’ of nucleosomes along the DNA by the class of ATP-hydrolyzing enzymatic machines called chromatin remodellers.We apply single pair-FRET for direct observation of intrinsic nucleosome dynamics as well as conformational changes of mononucleosomes induced by chromatin-remodeling complexes. The movement of DNA around the nucleosome surface is tracked in the presence and absence of remodeller by using mononucleosomes, which are reconstituted with a 200 bp long DNA containing the nucleosome positioning sequence 601 and a donor- and acceptor-dye pair at well-defined positions. Mononucleosomes are immobilized on DOPC-bilayer functionalized quartz slides and visualized by Total Internal Reflection Microscopy. |
| Databáze: | OpenAIRE |
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