Heterochromatic gene activation occurs in the context of minimal transcription-linked H3 methylation and is unimpaired by ablation of Dot1.

Abstract

<p>(a) H3K56ac ChIP analysis of <i>hsp82-2001</i> in <i>sir4Δ</i> or <i>SIR⁺</i> cells subjected to an instantaneous 30° to 39°C thermal upshift for the times indicated. Quantification was done using Real Time qPCR. The acetylated H3K56/Myc-H4 quotient of the non-induced <i>sir4Δ</i> sample was set to 1.0 for each amplicon. PTM-specific and Myc-H4 signals at the heat shock transgene were normalized to those measured at <i>PMA1</i> and <i>ARS504</i>, respectively. Shown are means ± S.D. (N = 2; qPCR = 4). (b) H3K36me3 ChIP analysis of <i>hsp82-2001</i> conducted as in A. (c) H3K4me3 ChIP analysis of <i>hsp82-2001</i> in <i>sir4Δ</i> or <i>SIR⁺</i> cells subjected to an instantaneous 30° to 39°C thermal upshift for the times indicated. Quantification and scaling were done as in A, except H3K4me3/H3 quotients are depicted, and both PTM-specific and H3 signals were normalized to those measured at <i>ARS504</i>. Shown are means ± S.D. (N = 2; qPCR = 4). (d) H3K79me2 ChIP analysis of <i>hsp82-2001</i> in <i>sir4Δ</i> or <i>SIR⁺</i> cells as in C. (e) Pol II ChIP analysis of heterochromatic <i>hsp82-2001</i> in <i>DOT1⁺</i> and <i>dot1Δ</i> strains subjected to heat shock as above. Pol II occupancy was determined using ChIP-qPCR as in <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1004202#pgen-1004202-g003" target="_blank">Figure 3A</a>. Shown are means ± S.D. (N = 2; qPCR = 4).</p