Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka, Japan.
PLoS One. 2020 Apr 3;15(4):e0231114. doi: 10.1371/journal.pone.0231114. eCollection 2020.
Meiosis and oocyte maturation are tightly regulated processes. The meiosis arrest female 1 (MARF1) gene is essential for meiotic progression in animals; however, its detailed function remains unclear. In this study, we examined the molecular mechanism of dMarf1, a Drosophila homolog of MARF1 encoding an OST and RNA Recognition Motif (RRM) -containing protein for meiotic progression and oocyte maturation. Although oogenesis progressed in females carrying a dMarf1 loss-of-function allele, the dMarf1 mutant oocytes were found to contain arrested meiotic spindles or disrupted microtubule structures, indicating that the transition from meiosis I to II was compromised in these oocytes. The expression of the full-length dMarf1 transgene, but none of the variants lacking the OST and RRM motifs or the 47 conserved C-terminal residues among insect groups, rescued the meiotic defect in dMarf1 mutant oocytes. Our results indicate that these conserved residues are important for dMarf1 function. Immunoprecipitation of Myc-dMarf1 revealed that several mRNAs are bound to dMarf1. Of those, the protein expression of nanos (nos), but not its mRNA, was affected in the absence of dMarf1. In the control, the expression of Nos protein became downregulated during the late stages of oogenesis, while it remained high in dMarf1 mutant oocytes. We propose that dMarf1 translationally represses nos by binding to its mRNA. Furthermore, the downregulation of Nos induces cycB expression, which in turn activates the CycB/Cdk1 complex at the onset of oocyte maturation.
减数分裂和卵母细胞成熟是受严格调控的过程。减数分裂停滞雌性 1 (MARF1)基因对于动物的减数分裂进展至关重要;然而,其详细功能仍不清楚。在这项研究中,我们研究了 dMarf1 的分子机制,dMarf1 是 MARF1 的果蝇同源物,编码一种含有 OST 和 RNA 识别基序(RRM)的蛋白质,对于减数分裂进展和卵母细胞成熟至关重要。尽管携带 dMarf1 功能丧失等位基因的雌性卵母细胞发生了卵发生,但发现 dMarf1 突变体卵母细胞中存在停滞的减数分裂纺锤体或微管结构破坏,表明这些卵母细胞中从减数分裂 I 向减数分裂 II 的转变受到了损害。全长 dMarf1 转基因的表达,但缺失 OST 和 RRM 基序或昆虫群中保守的 47 个 C 末端残基的任何变体都不能挽救 dMarf1 突变体卵母细胞的减数分裂缺陷。我们的结果表明这些保守残基对于 dMarf1 的功能很重要。Myc-dMarf1 的免疫沉淀表明有几种 mRNA 与 dMarf1 结合。在这些基因中,nos 的蛋白质表达(nos)受到影响,但不是其 mRNA 在没有 dMarf1 的情况下。在对照中,Nos 蛋白的表达在卵发生的后期阶段下调,而在 dMarf1 突变体卵母细胞中仍保持高水平。我们提出 dMarf1 通过与其 mRNA 结合来翻译抑制 nos。此外,Nos 的下调诱导 cycB 的表达,继而在卵母细胞成熟开始时激活 CycB/Cdk1 复合物。