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Effects of the loss of mismatch repair genes on single-strand annealing between divergent sequences in Saccharomyces cerevisiae

Journal of Microbiology 2021년 59권 4호 p.401 ~ 409
임예슬, 최주희, 안규진, 김민구, 배성호,
소속 상세정보
임예슬 ( Lim Ye-Seul ) - Inha University College of Natural Science Department of Biological Sciences
최주희 ( Choi Ju-Hee ) - Inha University College of Natural Science Department of Biological Sciences
안규진 ( Ahn Kyu-Jin ) - Inha University College of Natural Science Department of Biological Sciences
김민구 ( Kim Min-Ku ) - Inha University College of Natural Science Department of Biological Sciences
배성호 ( Bae Sung-Ho ) - Inha University College of Natural Science Department of Biological Sciences

Abstract


Eukaryotic genomes contain many duplicated genes closely located with each other, such as the hexose transporter (HXT) genes in Saccharomyces cerevisiae. They can potentially recombine via single-strand annealing (SSA) pathway. SSA between highly divergent sequences generates heteroduplex DNA intermediates with many mismatches, which can be corrected by mismatch repair (MMR), resulting in recombinant sequences with a single junction point. In this report, we demonstrate that SSA between HXT1 and HXT4 genes in MMR-deficient yeast cells produces recombinant genes with multiple-junctions resulting from alternating HXT1 and HXT4 tracts. The mutations in MMR genes had differential effects on SSA frequencies; msh6Δ mutation significantly stimulated SSA events, whereas msh2Δ and msh3Δ slightly suppressed it. We set up an assay that can identify a pair of recombinant genes derived from a single heteroduplex DNA. As a result, the recombinant genes with multiple-junctions were found to accompany genes with single-junctions. Based on the results presented here, a model was proposed to generate multiple-junctions in SSA pathway involving an alternative short-patch repair system.

키워드

single-strand annealing; mismatch repair; heteroduplex DNA; MSH2; MSH3; MSH6; hexose transporter gene; HO endonuclease; short-patch repair

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