Disruption of the intragenic transcription hampers chromatin remodeling and transcriptional initiation of ASP3. (A) Schematic diagram of the ASP3 locus. The three regions used for PCR amplification in the ChIP assay are marked below. (B–D) ChIP analyses performed at the ASP3 locus. Samples were collected at the indicated time points after nitrogen starvation. Regions 1–3 used for the PCR amplification are indicated. (B) ChIP analyses of histone H3 occupancy. (C) ChIP analyses of H3K4me3 at the ASP3 locus. (D) Rate of RNAPII-mediated initiation of transcription at the TATA-box of ASP3 was assayed by ChIP, using antibody against Ser5-phosphorylated CTD. Data are shown as relative levels compared to signals measured at time 0. (E) Nuclear run-on analysis was conducted to detect the transcription rate of ASP3 in the wild-type and iceΔ strains after nitrogen starvation. Fragments of ACT1 and the multiple cloning sites of the pBSKS plasmid were used as positive and negative controls, respectively. The ASP3 signals were normalized to that of ACT1 and are shown below.
