Instituto de Biología Funcional y Genómica (IBFG), CSIC-USAL, Salamanca, 37007, Spain.
Department of Genetics, Faculty of Natural Sciences, Comenius University in Bratislava, Bratislava, 841 04, Slovakia.
EMBO J. 2024 Oct;43(19):4356-4383. doi: 10.1038/s44318-024-00205-2. Epub 2024 Aug 22.
Meiosis is the developmental program that generates gametes. To produce healthy gametes, meiotic recombination creates reciprocal exchanges between each pair of homologous chromosomes that facilitate faithful chromosome segregation. Using fission yeast and biochemical, genetic, and cytological approaches, we have studied the role of CDK (cyclin-dependent kinase) in the control of Swi5-Sfr1, a Rad51-recombinase auxiliary factor involved in homolog invasion during recombination. We show that Sfr1 is a CDK target, and its phosphorylation downregulates Swi5-Sfr1 function in the meiotic prophase. Expression of a phospho-mimetic sfr1-7D mutant inhibits Rad51 binding, its robust chromosome loading, and subsequently decreases interhomolog recombination. On the other hand, the non-phosphorylatable sfr1-7A mutant alters Rad51 dynamics at late prophase, and exacerbates chromatin segregation defects and Rad51 retention observed in dbl2 deletion mutants when combined with them. We propose Sfr1 phospho-inhibition as a novel cell-cycle-dependent mechanism, which ensures timely resolution of recombination intermediates and successful chromosome distribution into the gametes. Furthermore, the N-terminal disordered part of Sfr1, an evolutionarily conserved feature, serves as a regulatory platform coordinating this phospho-regulation, protein localization and stability, with several CDK sites and regulatory sequences being conserved.
减数分裂是产生配子的发育程序。为了产生健康的配子,减数分裂重组在每对同源染色体之间创建相互交换,从而促进忠实的染色体分离。我们使用裂殖酵母和生化、遗传和细胞学方法,研究了 CDK(细胞周期蛋白依赖性激酶)在控制 Swi5-Sfr1 中的作用,Swi5-Sfr1 是一种参与重组过程中同源物入侵的 Rad51 重组辅助因子。我们表明 Sfr1 是 CDK 的靶标,其磷酸化下调了减数分裂前期 Swi5-Sfr1 的功能。表达磷酸模拟 sfr1-7D 突变体抑制 Rad51 结合,其强大的染色体加载,随后减少了同源重组。另一方面,非磷酸化 sfr1-7A 突变体改变了 Rad51 在后期的动力学,并与 dbl2 缺失突变体结合时加剧了染色质分离缺陷和 Rad51 的保留。我们提出 Sfr1 的磷酸化抑制是一种新的细胞周期依赖性机制,它确保重组中间体的及时解决,并成功将染色体分配到配子中。此外,Sfr1 的 N 端无序部分是一种进化保守的特征,作为一个调节平台,协调这种磷酸化调节、蛋白质定位和稳定性,几个 CDK 位点和调节序列被保守。