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A selection-based next generation sequencing approach to develop robust, genotype-specific mutation profiles in
Natalie A Lamb1, Jonathan E Bard2,3, Michael J Buck1,3
1Department of Biochemistry, Jacobs School of Medicine and Biomedical Sciences, University at Buffalo (SUNY), Buffalo, NY 14203, USA.
G3 (Bethesda, Md.)
|March 30, 2021
Summary
Altered deoxyribonucleotide triphosphate (dNTP) pools impact DNA replication fidelity. Our study reveals genotype-specific mutation profiles, linking dNTP pool changes to mutagenesis mechanisms.
Area of Science:
- Molecular Biology
- Genetics
- Biochemistry
Background:
- Mutation signatures can predict mutagenesis mechanisms, but altered deoxyribonucleotide triphosphate (dNTP) pools complicate these predictions.
- dNTP pool alterations impact DNA replication fidelity and overall mutation rates.
Purpose of the Study:
- To define mutation profiles associated with distinct rnr1 backgrounds, which affect dNTP pools.
- To investigate how altered dNTP pools influence replication fidelity and mutagenesis.
Main Methods:
- Targeted deep-sequencing of the CAN1 gene in Saccharomyces cerevisiae.
- Computational analysis of mutation spectra, position effects, and sequence context in different rnr1 mutant strains.
Main Results:
- Identified genotype-specific mutation profiles linked to altered dNTP pools, even with modest changes.
- Revealed sequence contexts and nucleotide motifs influencing variant profiles in different rnr1 backgrounds.
- Provided nuanced computational analysis of variants due to high sequencing depth.
Conclusions:
- Altered dNTP pools significantly impact replication fidelity and generate distinct mutation signatures.
- Mechanistic predictions based on mutation signatures must consider dNTP pool dynamics.
- This approach offers a robust method for understanding mutagenesis mechanisms related to dNTP pool alterations.
