The influence of sequence divergence between alleles of the human MS205 minisatellite incorporated into the yeast genome on length-mutation rates and lethal recombination events during meiosis

Certain minisatellites exhibit hypervariability with respect to the number of repeat units and, thus, allele length. Such polymorphism is generated by germline-specific recombinational events that occur at high frequencies and lead to the gain or loss of repeat units. In order to elucidate the molecular details of mutagenesis in minisatellites, we have integrated human minisatellites into the yeast genome in the vicinity of a hotspot for meiotic double-strand breaks (DSBs). Here, we describe the results of tetrad analyses of mutations in the human MS205 minisatellite in yeast strains heterozygous for alleles composed of 51 and 31 repeat units, as well as in a strain homozygous for the same 51 repeat unit allele. The length-mutation rate was twice as high in the heterozygous strain as in the homozygous strain, suggesting that sequence divergence between alleles enhances the generation of length mutations. In the case of heterozygotes, the frequency of length mutants resulting from inter-allelic exchange was significantly higher in tetrads with three viable spores than in tetrads with four viable spores, indicating that there is a higher probability for spore mortality in tetrads originating from meioses during which inter-allelic exchange of repeat units occurs. In an attempt to explain these findings, we propose a model for minisatellite mutation involving recombination, in which sequence divergence between alleles results in a heteroduplex containing numerous mismatches. We suggest that convergent mismatch-repair tracts in this heteroduplex give rise to a DSB that may be repaired by an additional round of recombination resulting in mutation of a third allele, or be lethal if such recombination fails. It appears probable that the formation of such additional mutants is the major explanation for the difference in meiotic length-mutation rates between the heterozygous and homozygous yeast strains, and that this phenomenon contributes to high germline length-mutation frequencies at minisatellites in humans.