IDENTIFICATION AND CHARACTERIZATION OF CEN12 IN THE BUDDING YEAST SACCHAROMYCES-CEREVISIAE

In this paper we report the cloning, sequencing and functional charact erization of CEN12 and an associated autonomously replicating sequence (ARS) from the budding yeast Saccharomyces cerevisiae. In the course of studying a dynamin-related gene, DNM1, we previously physically map ped the gene to chromosome 12. Genetic mapping showed that the gene wa s tightly linked (0.35 cM) to the centromere, Subcloning experiments r evealed that a centromere-like activity was included in a small segmen t of DNA immediately downstream from the DNM1 gene, Mitotic centromere activity was discerned by the ability of the region to de-stabilize a centromere-containing plasmid, and to stabilize an ARS-containing pla smid. Meiotic centromere activity was determined by the first-division segregation in crosses of ARS plasmids containing this region, The DN A sequence of this region revealed a sequence with strong homology to the consensus for yeast centromeres.