Analysis of cohesin-dependent gene regulation in fission yeast

In addition to its well-know function in chromosome segregation, increasing evidence implicates cohesin in the control of gene expression. It has been previously reported that inactivation of the cohesin loader Mis4 in G1-arrested cells leads to the dissociation of cohesin from chromatin. We exploited this experimental situation to ask whether this loss of cohesin would affect gene expression on a genome-wide scale. Three independent inoculates (biological replicates) of cells carring either the thermosensitive mis4-367 or the wild-type mis4+ allele were grown at permissive temperature of 25°C. Cells were G1-arreted by titrating out the Cdc10 transcription factor by overexpression of the C-terminal fragment of its binding partner Res1, and then shifted for 2 hours at 37°C. Cells were harvested before and after the temperature shift to proceed to total RNA extraction.