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High Throughput Sequencing Reveals Alterations in the Recombination Signatures with Diminishing Spo11 Activity

By Beth Rockmill (274696), Philippe Lefrançois (102385), Karen Voelkel-Meiman (129315), Ashwini Oke (199479), G. Shirleen Roeder (102392) and Jennifer C. Fung (199481)

Abstract

<div><p>Spo11 is the topoisomerase-like enzyme responsible for the induction of the meiosis-specific double strand breaks (DSBs), which initiates the recombination events responsible for proper chromosome segregation. Nineteen PCR-induced alleles of <i>SPO11</i> were identified and characterized genetically and cytologically. Recombination, spore viability and synaptonemal complex (SC) formation were decreased to varying extents in these mutants. Arrest by <i>ndt80</i> restored these events in two severe hypomorphic mutants, suggesting that <i>ndt80</i>-arrested nuclei are capable of extended DSB activity. While crossing-over, spore viability and synaptonemal complex (SC) formation defects correlated, the extent of such defects was not predictive of the level of heteroallelic gene conversions (prototrophs) exhibited by each mutant. High throughput sequencing of tetrads from <i>spo11</i> hypomorphs revealed that gene conversion tracts associated with COs are significantly longer and gene conversion tracts unassociated with COs are significantly shorter than in wild type. By modeling the extent of these tract changes, we could account for the discrepancy in genetic measurements of prototrophy and crossover association. These findings provide an explanation for the unexpectedly low prototroph levels exhibited by <i>spo11</i> hypomorphs and have important implications for genetic studies that assume an unbiased recovery of prototrophs, such as measurements of CO homeostasis. Our genetic and physical data support previous observations of DSB-limited meioses, in which COs are disproportionally maintained over NCOs (CO homeostasis).</p></div

Topics: Biological Sciences, throughput, sequencing, reveals, alterations, recombination, signatures, diminishing, spo11
Year: 2013
DOI identifier: 10.1371/journal.pgen.1003932
OAI identifier: oai:figshare.com:article/838467
Provided by: FigShare
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