Pol III transcription machinery occupancy monitoring upon rapamycin treatment

Effects of Maf1 deletion on the promoter occupancy during transcriptional repression by rapamycin Keywords: ChIP-Chip time course Epitope tagged chromatin proteins, Brf1-13myc and Bdp1-13myc (TFIIIB subunits), t95-13myc (TFIIIC subunit) and 3HA-C160 (Pol III subunit) were crosslinked to their targets in vivo. Cells were grown in rich YPD medium. Protein-DNA interactions were fixed in exponential growth phase without treatment or after 30 min of treatment with rapamycin. Genome-wide analysis of the enrichment of DNA fragments, immunopurified with antibodies directed to epitope tags, was performed in triplicate in each case. Enrichment ratio in replicates samples were normalized to the GeneList1 (median = 1) representing loci not targeted by the Pol III transcription machinery according to a previous study (Harismendy et al. 2003).