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.

Evolution: a collection of misfits.

Different strains of one genetic model species after another are turning out to have limited abilities to interbreed, as if they were on the way to becoming different species. Are model organisms aberrations, or are the first steps in speciation easier than they seem?

Rapid evolution of cheating mitochondrial genomes in small yeast populations.

Outcrossed sex exposes genes to competition with their homologues, allowing alleles that transmit more often than their competitors to spread despite organismal fitness costs. Mitochondrial populations in species with biparental inheritance are thought to be especially susceptible to such cheaters because they lack strict transmission rules like meiosis or maternal inheritance. Yet the interaction between mutation and natural selection in the evolution of cheating mitochondrial genomes has not been tested experimentally. Using yeast experimental populations, we show that although cheaters were rare in a large sample of spontaneous respiratory-deficient mitochondrial mutations (petites), cheaters evolve under experimentally enforced outcrossing even when mutation supply and selection are restricted by repeatedly bottlenecking populations.

Life-history evolution and density-dependent growth in experimental populations of yeast.

We studied the evolution of the correlation between growth rate r and yield K in experimental lineages of the yeast Saccharomyces cerevisiae. First, we isolated a single clone every approximately 250 generations from each of eight populations selected in a glucose-limited medium for 5000 generations at approximately 6.6 population doublings per day (20 clones per line × 8 lines) and measured its growth rate and yield in a new, galactose-limited medium (with ∼1.3 doubling per day). For most lines, r on galactose increased throughout the 5000 generations of selection on glucose whereas K on galactose declined. Next, we selected these 160 glucose-adapted clones in the galactose environment for approximately 120 generations and measured changes in r and K in galactose. In general, growth rate increased and yield declined, and clones that initially grew slowly on galactose improved more than did faster clones. We found a negative correlation between r and K among clones both within each line and across all clones. We provide evidence that this relationship is not heritable and is a negative environmental correlation rather than a genetic trade-off.

The yield of experimental yeast populations declines during selection.

The trade-off between growth rate and yield can limit population productivity. Here we tested for this life-history trade-off in replicate haploid and diploid populations of Saccharomyces cerevisiae propagated in glucose-limited medium in batch cultures for 5000 generations. The yield of single clones isolated from the haploid lineages, measured as both optical and population density at the end of a growth cycle, declined during selection and was negatively correlated with growth rate. Initially, diploid populations did not pay this cost of adaptation but haploidized after about 1000-3000 generations of selection, and this ploidy transition was associated with a decline in yield caused by reduced cell size. These results demonstrate the experimental evolution of a trade-off between growth rate and yield, caused by antagonistic pleiotropy, during adaptation in haploids and after an adaptive transition from diploidy to haploidy.

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.

Evolutionary genetics: desperate times call for more sex.

A new study has found that strains of the fungus Aspergillus nidulans produce more of their spores sexually in environments where they are less fit, resembling a hypothesized transitional stage in the evolution of sex.

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.

The role of sex in fungal evolution.

Sex in fungi is often associated with dispersal and dormancy, but in many species is not required for reproduction, and its evolutionary genetic role is unclear. Sex can accelerate adaptation to a new environment, and recombination in Saccharomyces cerevisiae, though historically rare, has had a prominent role in the origins of many sequenced strains, in particular the origin of clinical strains from domesticated ancestors. Sex and recombination have recently been discovered in several human pathogens that were long thought to be asexual, but so far there is no compelling evidence that it plays an important genetic role in their adaptation. The self-compatible (homothallic) sexual systems of many fungi severely limit their potential for recombination, but increase the rate at which they can segregate new adaptive mutations into homozygotes, a possible benefit of sex that has received much less attention than recombination.

Evolutionary genetics: a piggyback ride to adaptation and diversity.

Competition between adaptive mutations in different asexual lineages limits the rate of adaptation. But additional adaptive mutations can occur in lineages that already have one, altering the dynamics of evolving asexual populations.

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.

Experimental evolution with yeast.

Many of the difficulties of studying evolution in action can be surmounted using populations of microorganisms, such as yeast. A readily manipulated sexual system and an increasingly sophisticated array of molecular and genomic tools uniquely qualify Saccharomyces cerevisiae as an experimental subject. This minireview briefly describes some recent contributions of yeast experiments to current understanding of the evolution of ploidy, sex, mutation, and speciation.

Evolutionary genetics: choosing to evolve.

The evolution of mate choice is believed to be important in speciation. A recent experiment involving mating preference evolution in laboratory yeast populations supports theoretical predictions that this can occur without complete genetic isolation between populations, strengthening the case that ecological specialization as well as physical separation can lead to speciation.

Nuclear-mitochondrial epistasis for fitness in Saccharomyces cerevisiae.

In addition to the familiar possibility of epistasis between nuclear loci, interactions may evolve between the mitochondrial and nuclear genomes in eukaryotic cells. We looked for such interactions in Saccharomyces cerevisiae genotypes evolved independently and asexually in the laboratory for 2000 generations, and in an ecologically distinct pathogenic S. cerevisiae strain. From these strains we constructed derivatives entirely lacking mitochondrial DNA and then used crosses to construct matched and unmatched pairings of nuclear and mitochondrial genomes. We detected fitness effects of such interactions in an evolved laboratory strain and in crosses between the laboratory and pathogen strains. In both cases, there were significant contributions to progeny fitness of both nuclear and mitochondrial genomes and of their interaction. A second evolved genotype showed incompatibility with the first evolved genotype, but the nuclear and mitochondrial contributions to this incompatibility could not be resolved. These results indicate that cytonuclear interactions analogous to those already known from plants and animals can evolve rapidly on an evolutionary timescale.

The effects of sex and mutation rate on adaptation in test tubes and to mouse hosts by Saccharomyces cerevisiae.

Some hypotheses for the evolution of sex focus on adaptation to changing or heterogeneous environments, but these hypotheses have rarely been tested. We tested for advantages of sex and of increased mutation rates in yeast strains in two contrasting environments: a standard and relatively homogeneous laboratory environment of minimal medium in test tubes, and the variable environment of a mouse brain experienced by pathogenic strains. Evolving populations were founded as equal mixtures of sexual and obligately asexual genotypes. In the sexuals, cycles of sporulation, meiosis, and mating were induced approximately every 50 mitotic generations, with the asexuals undergoing sporulation but not ploidy cycles or recombination. In both environments, replicate negative control populations established with the same pair of strains were propagated with neither mating nor meiosis. In test tubes with no sex induced, sexuals were fixed in all five replicates within 250 mitotic generations, whereas in mice with no sex induced, asexuals were fixed in all four replicates by 170 generations. Inducing sex altered these outcomes in opposite directions in test tubes and mice, decreasing the fixation frequencies of sexuals in test tubes but increasing them in mice. These contrasts with asexual controls suggest an advantage for sex in mice but not in test tubes, although there was no difference between test tubes and mice in the numbers of populations fixed-for sexuals. In analogous experiments testing for an advantage of increased mutation rates, wild-type genotypes became fixed at the expense of mutators in every replicate of both test tube and mouse populations, indicating a disadvantage for mutators in both environments. Increased rates of point mutation do not appear to accelerate adaptation.

The number of mutations selected during adaptation in a laboratory population of Saccharomyces cerevisiae.

There is currently limited empirical and theoretical support for the prevailing view that adaptation typically results from the fixation of many mutations, each with small phenotypic effects. Recent theoretical work suggests that, on the contrary, most of the phenotypic change during an episode of adaptation can result from the selection of a few mutations with relatively large effects. I studied the genetics of adaptation by populations of budding yeast to a culture regime of daily hundredfold dilution and transfer in a glucose-limited minimal liquid medium. A single haploid genotype isolated after 2000 generations showed a 76% fitness increase over its ancestor. This evolved haploid was crossed with its ancestor, and tetrad dissections were used to isolate a complete series of six meiotic tetrads. The Castle-Wright estimator of the number of loci at which adaptive mutations had been selected, modified to account for linkage and variation among mutations in the size of their effect, is 4.4. The estimate for a second haploid genotype, isolated from a separate population and with a fitness gain of 60%, was 2.7 loci. Backcrosses to the ancestor with the first evolved genotype support the inference that adaptation resulted primarily from two to five mutations. These backcrosses also indicated that deleterious mutations had hitchhiked with adaptive mutations in this evolved genotype.

Antagonism between sexual and natural selection in experimental populations of Saccharomyces cerevisiae.

Trade-offs between life-history components are a central concept of evolution and ecology. Sexual and natural selection seem particularly apt to impose antagonistic selective pressures. When sex is not integrated into reproduction, as in Saccharomyces cerevisiae, natural selection can impair or even eliminate it. In this study, a genetic trade-off between the sexual and asexual phases of the yeast life cycle was suggested by sharp declines in the mating and sporulation abilities of unrelated genotypes that were propagated asexually in minimal growth medium and in mice. When sexual selection was applied to populations that had previously evolved asexually, sexual fitness increased but asexual fitness declined. No such negative correlation was observed when sexual selection was applied to an ancestral strain: sexual and asexual fitness both increased. Thus, evolutionary history affected the evolution of genetic correlations, as fitness increases in a population already well adapted to the environment were more likely to come at the expense of sexual functions.

Capturing the adaptive mutation in yeast.

An accurate view of adaptive mutations is essential to evolutionary genetics, but their rarity makes them difficult to study. This can be partially overcome using the many tools of yeast genetics and the ability to study very large populations over many generations. Adaptation to laboratory environments has occurred primarily by chromosomal rearrangements, often involving retrotransposons and apparently selected for their effects on gene regulation. Estimated rates of adaptive mutation are on the order of 1 in 10(11) cell divisions. There remains great potential for the genomic study of variation within yeast species to contribute to our understanding of adaptive mutation.