Evidence suggesting that the ARS elements associated with silencers of the yeast mating-type locus HML do not function as chromosomal DNA replication origins.
The silent mating-type loci of Saccharomyces cerevisiae, HML and HMR, are flanked by transcriptional silencers that have ARS activity (i.e., they function as replication origins when in plasmids). To test whether these ARS elements are chromosomal origins, we mapped origins near HML (close to the left telomere of chromosome III). Our results indicate that the HML-associated ARS elements either do not function as chromosomal replication origins or do so at a frequency below our detection level, suggesting that replication from a silencer-associated origin in each S phase is not essential for the maintenance of transcriptional repression at HML. Our results also imply that the ability of a DNA fragment to function as an ARS element in a plasmid does not ensure its ability to function as an efficient chromosomal replication origin. Telomere proximity is not responsible for inactivating these ARS elements, because they are not detectably functional as chromosomal origins even in genetically modified strains in which they are far from the telomere.
Dubey
D D
DD
Department of Molecular and Cellular Biology, Roswell Park Cancer Institute, Buffalo, New York 14263.
Davis
L R
LR
Greenfeder
S A
SA
Ong
L Y
LY
Zhu
J G
JG
Broach
J R
JR
Newlon
C S
CS
Huberman
J A
JA
D002874:Chromosome Mapping; D015825:Chromosomes, Fungal / Q000502:physiology*; D004261:DNA Replication / Q000502:physiology*; D015180:Electrophoresis, Gel, Two-Dimensional; D015966:Gene Expression Regulation, Fungal / Q000502:physiology; D005800:Genes, Fungal / Q000235:genetics*; D049770:Genes, Mating Type, Fungal; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics; D012093:Replicon / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics*