Journal of Microbiology

Ju-Hee Choi

2 Articles

Effects of the loss of mismatch repair genes on single-strand annealing between divergent sequences in Saccharomyces cerevisiae: Ye-Seul Lim , Ju-Hee Choi , Kyu-Jin Ahn , Min-Ku Kim , Sung-Ho Bae; J. Microbiol. 2021;59(4):401-409. Published online March 29, 2021; DOI: https://doi.org/10.1007/s12275-021-1076-x

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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.

Analyses of DNA double-strand break repair pathways in tandem arrays of HXT genes of Saccharomyces cerevisiae: Ju-Hee Choi , Ye-Seul Lim , Min-Ku Kim , Sung-Ho Bae; J. Microbiol. 2020;58(11):957-966. Published online October 30, 2020; DOI: https://doi.org/10.1007/s12275-020-0461-1

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Eukaryotic genomes contain numerous homologous repeat sequences including redundant genes with divergent homology that can be potential recombination targets. Recombination between divergent sequences is rare but poses a substantial threat to genome stability. The hexose transporter (HXT) gene family shares high sequence similarities at both protein and DNA levels, and some members are placed close together in tandem arrays. In this study, we show that spontaneous interstitial deletions occur at significantly high rates in HXT gene clusters, resulting in chimeric HXT sequences that contain a single junction point. We also observed that DNA double-strand breaks created between HXT genes produce primarily interstitial deletions, whereas internal cleavage of the HXT gene resulted in gene conversions as well as deletion products. Interestingly, interstitial deletions were less constrained by sequence divergence than gene conversion. Moreover, recombination-defective mutations differentially affected the survival frequency. Mutations that impair single-strand annealing (SSA) pathway greatly reduced the survival frequency by 10–1,000-fold, whereas disruption of Rad51-dependent homologous recombination exhibited only modest reduction. Our results indicate that recombination in the tandemly repeated HXT genes occurs primarily via SSA pathway.

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Deletion of IRC19 Causes Defects in DNA Double-Strand Break Repair Pathways in Saccharomyces cerevisiae
Ju-Hee Choi, Oyungoo Bayarmagnai, Sung-Ho Bae
Journal of Microbiology.2024; 62(9): 749. CrossRef
A novel CRISPR/Cas9 system with high genomic editing efficiency and recyclable auxotrophic selective marker for multiple-step metabolic rewriting in Pichia pastoris
Xiang Wang, Yi Li, Zhehao Jin, Xiangjian Liu, Xiang Gao, Shuyuan Guo, Tao Yu
Synthetic and Systems Biotechnology.2023; 8(3): 445. CrossRef
Enhancing Homologous Recombination Efficiency in Pichia pastoris for Multiplex Genome Integration Using Short Homology Arms
Jucan Gao, Cuifang Ye, Jintao Cheng, Lihong Jiang, Xinghao Yuan, Jiazhang Lian
ACS Synthetic Biology.2022; 11(2): 547. CrossRef
Effects of the loss of mismatch repair genes on single-strand annealing between divergent sequences in Saccharomyces cerevisiae
Ye-Seul Lim, Ju-Hee Choi, Kyu-Jin Ahn, Min-Ku Kim, Sung-Ho Bae
Journal of Microbiology.2021; 59(4): 401. CrossRef

Ju-Hee Choi

3 Articles

Erratum: Effects of the Loss of Mismatch Repair Genes on Single-Strand Annealing Between Divergent Sequences in Saccharomyces cerevisiae: Ye-Seul Lim, Ju-Hee Choi, Kyu-Jin Ahn, Min-Ku Kim, Sung-Ho Bae; J. Microbiol. 2024;62(10):929-929. Published online August 26, 2024; DOI: https://doi.org/10.1007/s12275-024-00126-z

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Erratum: Analyses of DNA Double-Strand Break Repair Pathways in Tandem Arrays of HXT Genes of Saccharomyces Cerevisiae: Ju-Hee Choi, Ye-Seul Lim, Min-Ku Kim, Sung-Ho Bae; J. Microbiol. 2024;62(10):931-931. Published online August 26, 2024; DOI: https://doi.org/10.1007/s12275-024-00127-y

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Deletion of IRC19 Causes Defects in DNA Double-Strand Break Repair Pathways in Saccharomyces cerevisiae: Ju-Hee Choi, Oyungoo Bayarmagnai, Sung-Ho Bae; J. Microbiol. 2024;62(9):749-758. Published online July 12, 2024; DOI: https://doi.org/10.1007/s12275-024-00152-x

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Abstract: DNA double-strand break (DSB) repair is a fundamental cellular process crucial for maintaining genome stability, with homologous recombination and non-homologous end joining as the primary mechanisms, and various alternative pathways such as single-strand annealing (SSA) and microhomology-mediated end joining also playing significant roles under specific conditions. IRC genes were previously identified as part of a group of genes associated with increased levels of Rad52 foci in Saccharomyces cerevisiae. In this study, we investigated the effects of IRC gene mutations on DSB repair, focusing on uncharacterized IRC10, 19, 21, 22, 23, and 24. Gene conversion (GC) assay revealed that irc10Δ, 22Δ, 23Δ, and 24Δ mutants displayed modest increases in GC frequencies, while irc19Δ and irc21Δ mutants exhibited significant reductions. Further investigation revealed that deletion mutations in URA3 were not generated in irc19Δ mutant cells following HO-induced DSBs. Additionally, irc19Δ significantly reduced frequency of SSA, and a synergistic interaction between irc19Δ and rad52Δ was observed in DSB repair via SSA. Assays to determine the choice of DSB repair pathways indicated that Irc19 is necessary for generating both GC and deletion products. Overall, these results suggest a potential role of Irc19 in DSB repair pathways, particularly in end resection process.

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