Mouse REC114 is essential for meiotic DNA double-strand break formation and forms a complex with MEI4
Mouse REC114 is essential for meiotic DNA double-strand break formation and forms a complex with MEI4
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DOI：
10.26508/lsa.201800259
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发表时间：
2018-12-01
影响因子：
4.4
通讯作者：
de Massy, Bernard
中科院分区：
文献类型：
--
作者：
Kumar, Rajeev;Oliver, Cecilia;de Massy, Bernard
DNA双链断裂(DSB)的程序性形成启动了减数分裂同源重组途径。这条途径对于第一次减数分裂时适当的染色体分离和生育是必不可少的。减数分裂DSB是由Spo11催化的。其他几种蛋白质对减数分裂DSB的形成也是必不可少的，包括三种在进化上首次在酿酒酵母中发现的保守蛋白质(Mer2，Mei4和Rec114)。这三种酿酒酵母蛋白及其小鼠同源基因(IHO1、MEI4和REC114)共定位于减数分裂染色体的轴上，而小鼠IHO1和MEI4是减数分裂DSB形成所必需的。在这里，我们证明了小鼠Rec114是减数分裂DSB形成所必需的。此外，MEI4与REC114和IHO1在小鼠精母细胞中形成了一个复合体，这与细胞学观察一致。然后，我们在体外证明了REC114 C末端结构域和MEI4 N末端结构域之间形成了稳定的复合体。我们进一步确定了REC114 N-末端结构域的结构，该结构显示了与Pleckstrin同源结构域的相似性。这些分析提供了对减数分裂DSB机制的这些基本组件的体系结构的直接见解。
Programmed formation of DNA double-strand breaks (DSBs) initiates the meiotic homologous recombination pathway. This pathway is essential for proper chromosome segregation at the first meiotic division and fertility. Meiotic DSBs are catalyzed by Spo11. Several other proteins are essential for meiotic DSB formation, including three evolutionarily conserved proteins first identified in Saccharomyces cerevisiae (Mer2, Mei4, and Rec114). These three S. cerevisiae proteins and their mouse orthologs (IHO1, MEI4, and REC114) co-localize on the axes of meiotic chromosomes, and mouse IHO1 and MEI4 are essential for meiotic DSB formation. Here, we show that mouse Rec114 is required for meiotic DSB formation. Moreover, MEI4 forms a complex with REC114 and IHO1 in mouse spermatocytes, consistent with cytological observations. We then demonstrated in vitro the formation of a stable complex between REC114 C-terminal domain and MEI4 N-terminal domain. We further determine the structure of the REC114 N-terminal domain that revealed similarity with Pleckstrin homology domains. These analyses provide direct insights into the architecture of these essential components of the meiotic DSB machinery.