The Kinetochore Is an Enhancer of Pericentric Cohesin Binding (original) (raw)
Figure 9
The Centromeric Enhancer Is Active at an Ectopic Location
The endogenous centromere on CHRIII was removed, and CEN6 was inserted at an ectopic location (SGD coordinate approximately 260 kb), producing yeast strain PMY318, as described in Materials and Methods. Cells containing the ectopic centromere and isogenic wild-type cells (1829-15B) were staged in G1 using αF, and then released into fresh medium containing nocodazole to arrest cells in mitosis. Cells were then fixed in formaldehyde and processed for ChIP using epitope-tagged Mcd1-6HAp as a marker for the cohesin complex.
(A) The Mcd1p binding profiles at the ectopic location on endogenous CHRIII (black squares) and in the presence of the ectopic centromere (gray circles) are shown. The location of the ectopic centromere is indicated by the black oval.
(B) The levels of Mcd1p binding in the region flanking the ectopically placed centromere were divided by those observed in the isogenic wild-type control strain to determine the fold increases in Mcd1p binding in the presence of the centromere. Data are plotted as a function of the SGD coordinates for this region.