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Genome-wide model for the normal eukaryotic DNA replication fork.

ABSTRACT: To investigate DNA replication enzymology across the nuclear genome of budding yeast, deep sequencing was used to establish the pattern of uncorrected replication errors generated by an asymmetric mutator variant of DNA polymerase ? (Pol ?). Sequencing of 16 genomes identified 1,206-bp substitutions generated over 33 generations by L612M Pol ? in a mismatch repair defective strain. Alignment of sequences flanking these substitutions identified "hotspot" motifs for Pol ? replication errors. The substitutions were distributed evenly across all 16 chromosomes. The vast majority were transitions that occurred with a strand bias that varied in a predictable manner relative to known functional origins of replication. This strand bias strongly supports the idea that Pol ? is primarily a lagging strand polymerase during replication across the entire nuclear genome.

SUBMITTER: Larrea AA

PROVIDER: S-EPMC2955150 | biostudies-literature | 2010 Oct

REPOSITORIES: biostudies-literature

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Genome-wide model for the normal eukaryotic DNA replication fork.

Larrea Andres A AA Lujan Scott A SA Nick McElhinny Stephanie A SA Mieczkowski Piotr A PA Resnick Michael A MA Gordenin Dmitry A DA Kunkel Thomas A TA

Proceedings of the National Academy of Sciences of the United States of America 20100927 41

To investigate DNA replication enzymology across the nuclear genome of budding yeast, deep sequencing was used to establish the pattern of uncorrected replication errors generated by an asymmetric mutator variant of DNA polymerase δ (Pol δ). Sequencing of 16 genomes identified 1,206-bp substitutions generated over 33 generations by L612M Pol δ in a mismatch repair defective strain. Alignment of sequences flanking these substitutions identified "hotspot" motifs for Pol δ replication errors. The s ...[more]

PMID: 20876092

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