Role for PADI6 in securing the mRNA-MSY2 complex to the oocyte cytoplasmic lattices
Role for PADI6 in securing the mRNA-MSY2 complex to the oocyte cytoplasmic lattices
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PADI6 在将 mRNA-MSY2 复合物固定到卵母细胞胞质晶格中的作用
DOI：
10.1080/15384101.2016.1261225
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发表时间：
2017-01-01
期刊：
影响因子：
4.3
通讯作者：
Coonrod, Scott
中科院分区：
文献类型：
--
作者：
Liu, Xiaoqiu;Morency, Eric;Coonrod, Scott
摘要卵母细胞胞质格（CPL）长期以来一直被认为是母体核糖体对早期胚胎贡献的一种储存形式。我们以前的研究表明，核糖体组分S6储存在卵母细胞CPL中，肽基精氨酸脱亚胺酶6（PADI 6）是CPL形成的关键。此外，我们发现PADI 6的缺失减少了母体到胚胎过渡之前的从头蛋白质合成，因此导致胚胎停滞在2细胞阶段。在这里，我们提出的证据进一步支持的关联核糖体与CPL证明，rRNA显着减少Padi 6 KO卵母细胞。我们还表明，mRNA的丰度和本地化的影响后，PADI 6耗尽，这表明mRNA是非常可能与CPL。与该观察结果一致，与Triton X-100提取后卵母细胞的不溶性部分相关的主要RNA结合蛋白MSY 2的量在Padi 6 KO卵母细胞中也显著降低。此外，用RNA酶A处理卵母细胞，然后用Triton X-100提取严重损害卵母细胞中PADI 6和MSY 2的定位。这些结果表明，可能与MSY 2和PADI 6形成复合物的mRNA结合在CPL中，并可能在将mRNA-MSY 2复合物固定到CPL中发挥作用。
ABSTRACT The oocyte cytoplasmic lattices (CPLs) have long been predicted to function as a storage form for the maternal contribution of ribosomes to the early embryo. Our previous studies have demonstrated that ribosomal component S6 is stored in the oocyte CPLs and peptidylarginine deiminase 6 (PADI6) is critical for CPLs formation. Additionally, we found that depletion of PADI6 reduced de novo protein synthesis prior to the maternal-to-embryonic transition, therefore causing embryos to arrest at the 2-cell stage. Here, we present evidence further supporting the association of ribosomes with the CPLs by demonstrating that rRNAs are dramatically decreased in Padi6 KO oocytes. We also show that the abundance and localization of mRNAs is affected upon PADI6 depletion, suggesting that mRNAs are very possibly associated with CPLs. Consistent with this observation, the amount of the major RNA binding protein, MSY2, that is associated with the insoluble fraction of the oocytes after Triton X-100 extraction is also markedly decreased in the Padi6 KO oocytes. Furthermore, treatment of the oocytes with RNase A followed by Triton X-100 extraction severely impairs the localization of PADI6 and MSY2 in oocytes. These results indicate that mRNAs, possibly in a complex with MSY2 and PADI6, are bound in the CPLs and may play a role in securing the mRNA-MSY2 complex to the CPLs.