Mutator Suppression and Escape from Replication Error–Induced Extinction in Yeast

Abstract

Cells rely on a network of conserved pathways to govern DNA replication fidelity. Loss of polymerase proofreading or mismatch repair elevates spontaneous mutation and facilitates cellular adaptation. However, double mutants are inviable, suggesting that extreme mutation rates exceed an error threshold. Here we combine alleles that affect DNA polymerase δ (Pol δ) proofreading and mismatch repair to define the maximal error rate in haploid yeast and to characterize genetic suppressors of mutator phenotypes. We show that populations tolerate mutation rates 1,000-fold above wild-type levels but collapse when the rate exceeds 10 ⁻³ inactivating mutations per gene per cell division. Variants that escape this error-induced extinction ( eex) rapidly emerge from mutator clones. One-third of the escape mutants result from second-site changes in Pol δ that suppress the proofreading-deficient phenotype, while two-thirds are extragenic. The structural locations of the Pol δ changes suggest multiple antimutator mechanisms. Our studies reveal the transient nature of eukaryotic mutators and show that mutator phenotypes are readily suppressed by genetic adaptation. This has implications for the role of mutator phenotypes in cancer.

Author Summary

Organisms strike a balance between genetic continuity and change. Most cells are well adapted to their niches and therefore invest heavily in mechanisms that maintain accurate DNA replication. When cell populations are confronted with changing environmental conditions, “mutator” clones with high mutation rates emerge and readily adapt to the new conditions by rapidly acquiring beneficial mutations. However, deleterious mutations also accumulate, raising the question: what level of mutational burden can cell populations sustain before collapsing? Here we experimentally determine the maximal mutation rate in haploid yeast. We observe that yeast can withstand a 1,000-fold increase in mutation rate without losing colony forming capacity. Yet no strains survive a 10,000-fold increase in mutation rate. Escape mutants with an “anti-mutator” phenotype frequently emerge from cell populations undergoing this error-induced extinction. The diversity of antimutator changes suggests that strong mutator phenotypes in nature may be inherently transient, ensuring that rapid adaptation is followed by genetic attenuation which preserves the beneficial, adaptive mutations. These observations are relevant to microbial populations during infection as well as the somatic evolution of cancer cells.