Co-activators and chromatin structure

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  • Co-Activators and Chromatin remodelling
    • ATP-dependent
    • 4 families
    • E.g. SWI/SNF - ubiquitous across eukaryotes - not always positive regulation
      • SWI/SNF remodelling - shown in DNase I footprinting studies
        • Add histones to DNA template - the position on the DNA will be the same in every experiment
        • DNase added, cuts in same way due to protection of DNA by histones - gives 10bp ladder
        • Addition of SWI/SNF + ATP --> disruption = further DNAse digestion
        • Gel shift experiments indicate nucleosomes are still present
        • SWI/SNF helps TBP to bind!
          • SWI/SNF alters nucleosomal structure facilitating binding of TBP
    • Can disrupt chromatin structure in many ways
      • Movement of nucleosomes relative to DNA template (sliding beads along string)
      • Eviction of octameric nucleosomes from DNA
        • NB. nucleosome = DNA wound around 8 histone protein cores
      • Eviction of individual histones
      • Exchange of histone proteins

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