Context dependent function of transcriptional regulator Rap1 in gene silencing and activation [ChIP-seq timecourse]
As part of a study on the function of Rap1 at HML and MAT in Saccharomyces cerevisiae, we performed an Anchor Away experiment on Rap1, inducibly depleting it from the nucleus upon addition of Rapamycin, followed by a ChIP-seq time-course. This was done in a strain that had SNPs introduced to HMLalpha to make it distinguishable from MATalpha. Samples were spiked with a constant amount (5% by OD) of a S.paradoxus strain that also expressed 2xV5-Rap1. Reads from the S.paradoxus sample served as the normalization factor. Peaks were then fit to a non-linear regression model to extract the k(off) and apparent residence time in vivo. Overall design: 2xV5-FRB-Rap1 ChIP-seq timecourse (t = 0 (DMSO), 5min, 10min, 15min, 20min, 30min, 45min, 60min) following addition of Rapamycin in S. cerevisiae  + Spike-in of 5% cells (by OD) of 2xV5-tagged S.paradoxus strain. Replicates are included for each experiment.