Division of labor at the eukaryotic replication fork.
DNA polymerase delta (Pol delta) and DNA polymerase epsilon (Pol epsilon) are both required for efficient replication of the nuclear genome, yet the division of labor between these enzymes has remained unclear for many years. Here we investigate the contribution of Pol delta to replication of the leading and lagging strand templates in Saccharomyces cerevisiae using a mutant Pol delta allele (pol3-L612M) whose error rate is higher for one mismatch (e.g., T x dGTP) than for its complement (A x dCTP). We find that strand-specific mutation rates strongly depend on the orientation of a reporter gene relative to an adjacent replication origin, in a manner implying that >90% of Pol delta replication is performed using the lagging strand template. When combined with recent evidence implicating Pol epsilon in leading strand replication, these data support a model of the replication fork wherein the leading and lagging strand templates are primarily copied by Pol epsilon and Pol delta, respectively.
Nick McElhinny
Stephanie A
SA
Laboratory of Molecular Genetics, National Institute of Environmental Health Sciences, NIH, DHHS, Research Triangle Park, NC 27709, USA.
Gordenin
Dmitry A
DA
Stith
Carrie M
CM
Burgers
Peter M J
PM
Kunkel
Thomas A
TA
D000483:Alleles; D053843:DNA Mismatch Repair; D004252:DNA Mutational Analysis; D004257:DNA Polymerase II / Q000235:genetics / Q000378:metabolism*; D004258:DNA Polymerase III; D004261:DNA Replication; D017930:Genes, Reporter; D008954:Models, Biological; D051718:MutS Homolog 2 Protein / Q000235:genetics / Q000378:metabolism; D009154:Mutation; D018741:Replication Origin; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*
C491232:POL3 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D004257:DNA Polymerase II; D004258:DNA Polymerase III; C489751:MSH2 protein, S cerevisiae; D051718:MutS Homolog 2 Protein