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. 2009 Dec 21;38(6):1805–1820. doi: 10.1093/nar/gkp1172

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© The Author(s) 2009. Published by Oxford University Press.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 2. — Effects of the taf1-T657K mutation on the localization of GTFs and pol II at a subset of class II gene promoters. Strains expressing HA-tagged TAF1 (YTK2741) or taf1-T657K (YTK3780) alone or in combination with PK-tagged GCN5 (YTK6831/YTK6858; this depicts TAF1/taf1-T657K strains, respectively), SUA7 (YTK6837/YTK6864), TFA2 (YTK6838/YTK6865), TFG1 (YTK6839/YTK6866) or TFB3 (YTK6840/YTK6867) were cultured and cross-linked as described in Figure 1. The cross-linked chromatin was prepared and precipitated with anti-CTD (Rpb1, H), anti-HA (Taf1, A) or anti-PK (Gcn5, B; Sua7, C–D; Tfa2, E; Tfg1, F; Tfb3, G) monoclonal antibodies. After the cross-link reversal, quantitative PCR was carried out in triplicate to determine the recovery ratio of DNA corresponding to the promoters of several genes or POL1 ORF (negative control) as indicated at the bottom of each panel. The average values from three independent experiments with standard deviations of the ratios of precipitated DNA to the inputs are shown as white (TAF1) or black (taf1-T657K) bars in each panel. YTK2741 and YTK3780 strains were used to assess occupancy levels of Taf1 and Rpb1. Note that the data for Sua7 are shown in two panels (C and D) with different scales on the vertical axis as occupancy levels varied greatly between PGK1 and ribosomal protein genes.