Abstract
Crossing-over is a reciprocal exchange of chromosome arms and is required for accurate segregation of homologs during the first meiotic division. Currently the mechanism of crossing-over is described by the Double- Strand Break Repair Model (DSBR), which predicts the formation of programmed double strand breaks that are modified to allow strand invasion of an intact homolog. Single-end invasions (SEI) and double Holliday junctions (dHJ) are intermediates of meiotic recombination which have been characterized only by indirect criteria. Here we assess Atomic Force Microscopy (AFM) as a potentially powerful tool to study dHJ and SEI intermediates in Saccharomyces cerevisiae. We hypothesize that SEIs are classical displacement-loop structures while dHJs comprise two exchange junctions, positioned symmetrically about the double strand break site. Analysis of joint molecules is performed using crude extract of joint molecules isolated from ndt80-Δ1 (meiotic arrest mutant) as well as at the well characterized HIS4LEU2 meiotic recombination hotspot.
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