Yu J, Hecht N B, Schultz R M
Department of Biology, University of Pennsylvania, Philadelphia, Pennsylvania 19104-6018, USA.
Biol Reprod. 2001 Oct;65(4):1260-70. doi: 10.1095/biolreprod65.4.1260.
Translational control plays a central role during oocyte maturation and early embryogenesis, as these processes occur in the absence of transcription. MSY2, a member of a multifunctional Y-box protein family, is implicated in repressing the translation of paternal mRNAs. Here, we characterize MSY2 expression in mouse oocytes and preimplantation embryos. Northern blot analysis indicates that MSY2 expression is highly restricted and essentially confined to the oocyte in the female mouse. MSY2 transcript and protein, as assessed by reverse transcription-polymerase chain reaction and immunoblotting, respectively, are expressed in growing oocytes, metaphase II-arrested eggs, and 1-cell embryos, but then are degraded by the late 2-cell stage; no expression is detectable in the blastocysts. During oocyte maturation, MSY2 is phosphorylated and following fertilization it is dephosphorylated. Quantification of the mass amount of MSY2 reveals that it represents 2% of the total protein in the fully grown oocyte, i.e., it is a very abundant protein. Both endogenous MSY2 and MSY2-enhanced green fluorescent protein (EGFP), which is synthesized following microinjection of an mRNA encoding MSY2-EGFP, are primarily localized in the cytoplasm, and about 75% of the MSY2 remains associated with oocyte cytoskeletal preparations. Results of these studies are consistent with the proposal that MSY2 functions by stabilizing and/or repressing the translation of maternal mRNAs.
翻译控制在卵母细胞成熟和早期胚胎发育过程中起着核心作用,因为这些过程是在没有转录的情况下发生的。MSY2是多功能Y盒蛋白家族的成员之一,与抑制父本mRNA的翻译有关。在这里,我们描述了MSY2在小鼠卵母细胞和植入前胚胎中的表达情况。Northern印迹分析表明,MSY2的表达受到高度限制,基本上局限于雌性小鼠的卵母细胞中。通过逆转录-聚合酶链反应和免疫印迹分别评估的MSY2转录本和蛋白,在生长中的卵母细胞、中期II期阻滞的卵和1细胞胚胎中表达,但在2细胞晚期被降解;在囊胚中未检测到表达。在卵母细胞成熟过程中,MSY2被磷酸化,受精后去磷酸化。对MSY2质量量的定量分析表明,它占完全成熟卵母细胞总蛋白的2%,即它是一种非常丰富的蛋白。内源性MSY2和通过显微注射编码MSY2-增强绿色荧光蛋白(EGFP)的mRNA后合成的MSY2-EGFP主要定位于细胞质中,约75%的MSY2仍与卵母细胞细胞骨架制剂相关。这些研究结果与MSY2通过稳定和/或抑制母本mRNA的翻译发挥作用的提议一致。