Fig. 2

Spt2, Spt6, and Spt16 have reduced occupancy over SRG1, a highly transcribed RNA, in histone mutations. ChIP of histone H3 (a), H2B (b), Spt6 (c), Spt16 (d), Spt2-13myc (f), HA-Paf1 (g), or Rpb3 (h) was performed on chromatin isolated from strains expressing HHTS-HHFS alleles (YS454-YS456) or the indicated histone mutant alleles (YS458-YS462, YS465, YS471, YS472, and YS474) that were grown in YPD at 30 °C. HHTS-HHFS alleles are a synthetic histone gene sequence previously developed [52] and replacing each the HHT1-HHF1 and HHT2-HHF2 alleles. ChIP of Asf1-TAP (e) was performed on chromatin prepared from strains expressing HHTS-HHFS alleles (YS493-YS495) or the indicated histone mutant alleles (YS504-YS506, YS518, YS519, YS521, YS525-YS527) that were grown in YPD at 30 °C. The amount of immunoprecipitated DNA was determined by qPCR and is shown as a percentage of the input material and represents the mean ± SEM of three biological replicate experiments. Below the graphs is a schematic of the SRG1/SER3 locus with black bars corresponding to the regions amplified by qPCR