Functional Centromeres Determine the Activation Time of Pericentric Origins of DNA Replication in Saccharomyces cerevisiae
(A) Z-score plot of chromosome XV in WT (black) and rearranged (blue) strains. ARS1531 displayed a difference of Z-score values at least as large as that seen for ARS1410 and ARS1426 (see Figure 5B). See Figure S4 for all chromosomes. (B) 2D gel analysis of ARS1531 in the WT (left), the rearranged strain used in microarray analysis (middle), and the rearranged strain used for prior slot blot analysis (right). DNA from all three strains was digested with NcoI and BglII to give a 3.18 kb fragment harboring ARS1531 and then subjected to 2D gel analysis. The presence of a bubble arc in the WT (black arrow) indicates that ARS1531 is a functional origin in this strain. The presence of a bubble arc in one of the two rearranged strains confirms that the absence of origin activity in rearranged A (used in microarray analysis) is not due to relocation of the centromere on chromosome XIV. Below each 2D gel image is the sequence for the WT or mutant (red) ACS.