| Popis: |
DNA in eukaryotes is highly compacted. However, it is essential that proteins can have controlled access to the DNA during a number of fundamental cellular processes such as replication, transcription or repair. A large family of protein complexes commonly referred to as Swi2/Snf2 complexes allows for the transient remodelling of the essential compaction unit, the nucleosome and thus assures the access to, or repression of certain segments of DNA. In spite of a large number of research efforts using bio-chemical, structural and theoretical approaches the molecular mechanism of the ATP dependent nucleosome remodelling is currently not well understood. We therefore performed single molecule FRET experiments aimed to unravel details of the remodelling kinetics and pathway.We present remodelling data obtained by the remodelling complex ACF from drosophila on mono-nucleosomal constructs that use the 601- localisation sequence in combination with a biochemically well characterised linker DNA. The remodelling data was obtained, both, on nucleosomes immobilised onto surfaces of micro-fluidic chambers as well as from solution measurements using pulsed interleaved excitation and multi-parameter fluorescence detection. |
