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Lecture 3

Chromatin Remodeling

QuestionAnswer
What is MNase digestion and mapping? positioning of nucleosomes relative to known sequence; small
What is DNase digestion and maping? access to the DNA on or near nucleosomes, distortions; bigger
What is Electrophoretic Mobility Shift Assay (EMSA)? DNA bound to protein will run more slowly in a non-denaturing PAGE gel
What is the SWI/SNF class of remodelers? large complexes containing actin proteins; cause sliding and ejection of histones; binds acetylated histones
What is the SWI/Snf function? required for gene expression; 80 bp at a time
What is the RSC function? required for kinetochore function and cohesion in the arms of the kinetochore.
What is the ISWI class of remodelers? large protein complex that makes small changes in nucleosome assembly and spacing; 10 bp at a time; binds unmodified histones with SANT
What is the CHD class of remodelers? provides a link between histone modification and covalent modification; binds methylated histones
What is the split ATPase class of remodelers? Ino80 and SWR1
What is the function of Ino80? repairs damaged DNA by phosphorylated histones
What is the function of SWR1? swaps variant histones into the nucleosomes of actively transcribed genes; H2AZ for H2A
Where does acetylation occur? lysine
Where does methylation occur? lysine and arginine
Where does phosphorylation occur? serine and threonine
Where does ubiquitylation occur? lysine
What is ubiquitin? signaling module; multiple targets protein for degradation; single functions as a switch
What is Western blot? SDS-PAGE protein separation; antibody detected by a chemically labeled secondary antibody
What is Immunoprecipitation (IP)? extract treated with antibody; antibody is isolated and bound to target protein
What is Chromatin Immunoprecipitation (ChIP)? isolates proteins with DNA associated
Created by: javaughnr
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