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Related Experiment Video

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Strand-Specific Analysis of Proteins at Replicating DNA Strands by Enrichment and Sequencing of Protein-Associated Nascent DNA Method

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

Andres A Larrea¹, Scott A Lujan, Stephanie A Nick McElhinny

¹Laboratory of Molecular Genetics and Laboratory of Structural Biology, Department of Health and Human Services, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, NC 27709, USA.

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

|September 30, 2010

Summary

This summary is machine-generated.

Deep sequencing revealed DNA polymerase δ (Pol δ) replication errors in yeast. These errors, primarily transitions, showed a strand bias supporting Pol δ

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Area of Science:

Molecular Biology
Genetics
Biochemistry

Background:

DNA polymerase δ (Pol δ) is a key enzyme in DNA replication.
Understanding replication fidelity is crucial for genome stability.

Purpose of the Study:

To investigate DNA replication enzymology using an asymmetric mutator variant of DNA polymerase δ (Pol δ).
To establish the pattern of uncorrected replication errors across the budding yeast nuclear genome.

Main Methods:

Deep sequencing of 16 yeast genomes.
Analysis of 1,206 base pair substitutions generated by L612M Pol δ in a mismatch repair defective strain over 33 generations.
Identification of "hotspot" motifs for Pol δ replication errors by aligning flanking sequences.

Main Results:

Substitutions were evenly distributed across all 16 chromosomes.
The majority of substitutions were transitions with a strand bias.
This strand bias correlated predictably with known functional origins of replication.

Conclusions:

The observed strand bias strongly supports DNA polymerase δ (Pol δ) acting as a lagging strand polymerase genome-wide.
This study provides insights into the replication enzymology and error patterns of Pol δ.