Department of Molecular Biology and Genetics, Cornell University, Ithaca, NY 14853, USA.
Genetics. 2024 Jul 8;227(3). doi: 10.1093/genetics/iyae066.
The accurate segregation of homologous chromosomes during the Meiosis I reductional division in most sexually reproducing eukaryotes requires crossing over between homologs. In baker's yeast approximately 80% of meiotic crossovers result from Mlh1-Mlh3 and Exo1 acting to resolve double-Holliday junction intermediates in a biased manner. Little is known about how Mlh1-Mlh3 is recruited to recombination intermediates to perform its role in crossover resolution. We performed a gene dosage screen in baker's yeast to identify novel genetic interactors with Mlh1-Mlh3. Specifically, we looked for genes whose lowered dosage reduced meiotic crossing over using sensitized mlh3 alleles that disrupt the stability of the Mlh1-Mlh3 complex and confer defects in mismatch repair but do not disrupt meiotic crossing over. To our surprise we identified genetic interactions between MLH3 and DMC1, the recombinase responsible for recombination between homologous chromosomes during meiosis. We then showed that Mlh3 physically interacts with Dmc1 in vitro and in vivo. Partial complementation of Mlh3 crossover functions was observed when MLH3 was expressed under the control of the CLB1 promoter (NDT80 regulon), suggesting that Mlh3 function can be provided late in meiotic prophase at some functional cost. A model for how Dmc1 could facilitate Mlh1-Mlh3's role in crossover resolution is presented.
在大多数有性繁殖的真核生物中,减数分裂 I 减数分裂过程中同源染色体的精确分离需要同源染色体之间的交叉。在面包酵母中,大约 80%的减数分裂交叉是由 Mlh1-Mlh3 和 Exo1 以偏向的方式作用于解决双 Holliday 连接中间体而产生的。关于 Mlh1-Mlh3 如何被招募到重组中间体以发挥其在交叉分辨率中的作用,我们知之甚少。我们在面包酵母中进行了基因剂量筛选,以鉴定与 Mlh1-Mlh3 具有新型遗传相互作用的基因。具体来说,我们寻找那些降低剂量会降低使用敏感 mlh3 等位基因的减数分裂交叉的基因,这些等位基因会破坏 Mlh1-Mlh3 复合物的稳定性并导致错配修复缺陷,但不会破坏减数分裂交叉。令我们惊讶的是,我们发现 MLH3 与 DMC1 之间存在遗传相互作用,DMC1 是减数分裂过程中同源染色体之间重组的重组酶。然后,我们表明 Mlh3 在体外和体内与 Dmc1 物理相互作用。当 MLH3 在 CLB1 启动子(NDT80 调节子)的控制下表达时,观察到 Mlh3 交叉功能的部分互补,这表明 Mlh3 功能可以在减数前期的某个功能成本下提供晚期。提出了 Dmc1 如何促进 Mlh1-Mlh3 在交叉分辨率中的作用的模型。