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Our study used various experimental approaches to examine the mutational mechanisms operating within mature microsatellites, which we define as alleles that are not expected to mutate to lengths below or at the threshold for microsatellite mutational behavior in one round of replication.
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We used our established in vitro HSV-tk mutagenesis assay (Messier et al. 1996; Eckert et al. 2002; Eckert and Hile 2009; Kelkar et al. 2010) to evaluate experimentally the specific contribution of polymerase strand slippage errors during DNA synthesis to the observed mutational behavior of TRs in the human genome.
The differences in mutational behavior between TR classes could be due, in part, to differences in the efficiency of cellular mismatch repair (MMR).
The change points in mutational behavior that we identify here correspond to previously defined MS threshold values.
Our work combines computational and experimental approaches to explain the complex mutational behavior of dinucleotide microsatellites in humans.
This length also corresponds to a transition in mutational behavior of intronic and exonic trinucleotide repeats, as noted by Molla et al. (2009).
In conclusion, we emphasize that none of the above models are mutually exclusive and that several mechanisms may cooperatively contribute to the dramatic change in mutational behavior that is observed upon increasing the length of a tandemly repeated sequence.
A second explanation for the length-dependent change in mutational behavior is that alternative, non-B-form DNA structures are stabilized within the repetitive array after the transition length has been reached.
This suggests that the change in mutational behavior depends on the total length of the repetitive array, corroborating some previous studies (Rose and Falush 1998; Dieringer and Schlötterer 2003).
Based on changes in mutational behavior, we have defined the threshold length at which a short tandem repeat becomes a microsatellite (Kelkar et al. 2010; Ananda et al. 2013).
The availability of sequenced human genomes from the Pilot 1 phase of the 1000 Genomes Project (The 1000 Genomes Project Consortium 2010) provides an excellent opportunity to study the incidence of TR polymorphism and changes in mutational behavior that occur on a genome-wide scale and across human populations.
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