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. Author manuscript; available in PMC: 2014 Sep 12.
Published in final edited form as: Cell. 2013 Sep 12;154(6):10.1016/j.cell.2013.08.005. doi: 10.1016/j.cell.2013.08.005

Figure 4. Swc2 is required for SWR1 targeting in vivo and binds DNA directly.

Figure 4

(A) ChIP analysis of Swr1-TAP. DNA hybridization to a tiling microarray covering chromosome 3 and 6. The graphs show geometric mean of SWR1 signal for 226 genes ±1 Kb around TSS for WT and mutant strains. Data shown twice for WT (Left and Center) and htz1Δ (Center and Right).

(B) Map of Swc2 protein. Basic (blue) and acidic (red) amino acids. Grey bars mark evolutionarily conserved domains in Swc2.

(C) EMSA showing binding of bacterially expressed Swc2 segments to free DNA.

(D) Affinity-purified (*) and glycerol gradient purified SWR1 complexes resolved by SDS-PAGE and stained with Coomassie Blue.

(E–F) EMSA showing binding of WT and mutant SWR1 complexes to Alexa 647 labeled DNA and Cy5 labeled mono-nucleosome.

(G) ChIP-PCR analysis of Swr1-TAP binding to FUN12 and SWR1 promoters in WT and swc2 (136–345Δ) mutant cells. Tel: sub-telomeric sequence on chr 6. In mutant cells, reduced Swr1-TAP binding was also seen at RPL4A, SNT1 and NUP1 gene promoters (data not shown).

See also Figure S3 and S4.