Loss of Elongation-Like Factor 1 Spontaneously Induces Diverse, RNase H-Related Suppressor Mutations in Schizosaccharomyces pombe.

A healthy individual may carry a detrimental genetic trait that is masked by another genetic mutation. Such suppressive genetic interactions, in which a mutant allele either partially or completely restores the fitness defect of a particular mutant, tend to occur between genes that have a confined functional connection. Here we investigate a self-recovery phenotype in Schizosaccharomyces pombe, mediated by suppressive genetic interactions that can be amplified during cell culture. Cells without Elf1, an AAA+ family ATPase, have severe growth defects initially, but quickly recover growth rates near to those of wild-type strains by acquiring suppressor mutations. elf1Δ cells accumulate RNAs within the nucleus and display effects of genome instability such as sensitivity to DNA damage, increased incidence of lagging chromosomes, and mini-chromosome loss. Notably, the rate of phenotypic recovery was further enhanced in elf1Δ cells when RNase H activities were abolished and significantly reduced upon overexpression of RNase H1, suggesting that loss of Elf1-related genome instability can be resolved by RNase H activities, likely through eliminating the potentially mutagenic DNA-RNA hybrids caused by RNA nuclear accumulation. Using whole genome sequencing, we mapped a few consistent suppressors of elf1Δ including mutated Cue2, Rpl2702, and SPBPJ4664.02, suggesting previously unknown functional connections between Elf1 and these proteins. Our findings describe a mechanism by which cells bearing mutations that cause fitness defects and genome instability may accelerate the fitness recovery of their population through quickly acquiring suppressors. We propose that this mechanism may be universally applicable to all microorganisms in large-population cultures.

A Potential Case of Reinforcement in a Facultatively Sexual Unicellular Eukaryote.

The origin of a new species requires a mechanism to prevent divergent populations from interbreeding. In the classic allopatric model, divided populations evolve independently and accumulate genetic differences. If contact is restored, hybrids suffer reduced fitness and selection may favor traits that prevent mistakes in mating, a process known as reinforcement. This decisive but transient phase is challenging to document and has been reported mostly in macroorganisms. Very little is known about the processes through which new microbial species originate. In particular, it is unclear whether microbial eukaryotes, many of which can reproduce sexually during complex life cycles, speciate in much the same way as do well-studied plants and animals. Using individual cellular mate choice trials, we investigated the mating behavior of sympatric and allopatric woodland populations of the yeast Saccharomyces paradoxus. We find evidence consistent with reinforcement, potentially representing an example of microbial speciation in progress.

Prezygotic isolation between Saccharomyces cerevisiae and Saccharomyces paradoxus through differences in mating speed and germination timing.

Although prezygotic isolation between sympatric populations of closely related animal and plant species is well documented, far less is known about such evolutionary phenomena in sexual microbial species, as most are difficult to culture and manipulate. Using the molecular and genetic tools available for the unicellular fungus Saccharomyces cerevisiae, and applying them to S. paradoxus, we tested the behavior of individual cells from sympatric woodland populations of both species for evidence of prezygotic isolation. First, we confirmed previous observations that vegetative cells of both species mate preferentially with S. cerevisiae. Next, we found evidence for mate discrimination in spores, the stage in which outcrossing opportunities are most likely to occur. There were significant differences in germination timing between the species: under the same conditions, S. paradoxus spores do not begin germinating until almost all S. cerevisiae spores have finished. When germination time was staggered, neither species discriminated against the other, suggesting that germination timing is responsible for the observed mate discrimination. Our results indicate that the mechanisms of allochronic isolation that are well known in plants and animals can also operate in sexual microbes.

Yeast sex: surprisingly high rates of outcrossing between asci.

BACKGROUND: Saccharomyces yeasts are an important model system in many areas of biological research. Very little is known about their ecology and evolution in the wild, but interest in this natural history is growing. Extensive work with lab strains in the last century uncovered the Saccharomyces life cycle. When nutrient limited, a diploid yeast cell will form four haploid spores encased in a protective outer layer called the ascus. Confinement within the ascus is thought to enforce mating between products of the same meiotic division, minimizing outcrossing in this stage of the life cycle. METHODOLOGY/PRINCIPAL FINDINGS: Using a set of S. cerevisiae and S. paradoxus strains isolated from woodlands in North America, we set up trials in which pairs of asci were placed in contact with one another and allowed to germinate. We observed outcrossing in approximately 40% of the trials, and multiple outcrossing events in trials with three asci in contact with each other. When entire populations of densely crowded asci germinated, approximately 10-25% of the resulting colonies were outcrossed. There were differences between the species with S. cerevisiae having an increased tendency to outcross in mass mating conditions. CONCLUSIONS/SIGNIFICANCE: Our results highlight the potential for random mating between spores in natural strains, even in the presence of asci. If this type of mating does occur in nature and it is between close relatives, then a great deal of mating behavior may be undetectable from genome sequences.

A short history of recombination in yeast.

Despite it being the darling of fungal genomics, we know little about either the ecology or reproductive biology of the budding yeast, Saccharomyces cerevisiae, in nature. A recent study by Ruderfer et al. estimated that the ancestors of three S. cerevisiae genomes outcrossed approximately once every 50,000 generations, confirming the view that outcrossing is infrequent in natural populations of S. cerevisiae. This study also inferred the genomic positions of past recombination events. By comparing past recombination events with present-day recombination rates, this study lays the groundwork for determining whether recombination has improved the long-term survival of descendant lineages by bringing together favorable alleles, a longstanding question in evolutionary genetics.

HETEROMORPHISM FOR A HIGHLY REPEATED SEQUENCE IN THE NEW ZEALAND FROG LEIOPELMA HOCHSTETTERI.

A satellite DNA sequence, Lhl, was cloned from the New Zealand endemic frog Leiopelma hochstetteri. Large tandem arrays of Lh1 were localized by in situ hybridization to the long arm of a small telocentric autosome in some individuals, but these arrays were absent from other individuals. Lh1 is also present in varying amounts on some supernumerary chromosomes in some individuals. Heteromorphism for the presence of Lh1 exists in two populations that have been separated by a sea channel since the end of the Pleistocene, indicating that the heteromorphism either has arisen repeatedly or has persisted for at least 10,000 years. Individuals lacking Lh1 thus appear to be at no significant selective disadvantage. The variation in Lh1 copy number probably results from its interstitial chromosomal location, which exposes it to more frequent unequal crossovers than the pericentromeric or telocentric locations of most satellite DNA. Lh1 may be parasitic or simply inert junk, but in either case it may be deleted or dispersed throughout the rest of the genome through unequal crossing over.