Loss of Cnot6l Impairs Inosine RNA Modifications in Mouse Oocytes.
Loss of Cnot6l Impairs Inosine RNA Modifications in Mouse Oocytes.
复制标题
DOI：
10.3390/ijms22031191
复制
发表时间：
2021-01-26
影响因子：
5.6
通讯作者：
Christenson LK
中科院分区：
文献类型：
--
作者：
Brachova P;Alvarez NS;Christenson LK
哺乳动物卵母细胞必须通过一个称为翻译mRNA衰变的过程降解母体转录物，其中母体mRNA经历翻译激活，然后是去腺苷化和mRNA衰变。一旦转录物被激活，它就会被CCR 4-NOT复合物去腺苷化。CCR 4-NOT转录复合物亚基6样（Cnot 6l）（CCR 4-NOT复合物内的去腺苷酶）的敲除导致中期I（MI）进入期间mRNA衰变缺陷。B细胞易位基因-4（Btg 4）是CCR 4-NOT复合物的一种衔接蛋白，敲除Btg 4会导致受精后mRNA衰变缺陷。因此，控制mRNA周转的机制对卵母细胞的能力和早期胚胎发育有重要影响。转录后肌苷RNA修饰可能通过翻译机制影响mRNA的稳定性。在这里，我们评估了来自Cnot 6l-/-和Btg 4-/-小鼠的卵母细胞、卵子和胚胎中的肌苷RNA修饰，这些小鼠显示mRNA的稳定化和稳定化转录物的过度翻译。如果肌苷修饰在调节RNA稳定性中起作用，我们假设在这些突变背景中，我们将观察到肌苷mRNA修饰的变化或破坏。为了测试这一点，我们使用计算方法来鉴定减数分裂成熟（GV、MI和MII阶段）期间总的和多聚体RNA-seq数据中的肌苷RNA修饰。我们在Cnot 6l-/-小鼠的样品中观察到肌苷mRNA修饰的显著耗竭，但在Btg 4-/-小鼠中没有。此外，核糖体相关RNA的分析显示肌苷修饰的mRNA的清除。这些观察结果表明，在卵母细胞成熟过程中的mRNA清除的一种新的机制，其中含肌苷的成绩单衰减在一个独立的，但平行的机制CCR 4-NOT去腺苷化。
Mammalian oocytes must degrade maternal transcripts through a process called translational mRNA decay, in which maternal mRNA undergoes translational activation, followed by deadenylation and mRNA decay. Once a transcript is translationally activated, it becomes deadenylated by the CCR4-NOT complex. Knockout of CCR4-NOT Transcription Complex Subunit 6 Like (Cnot6l), a deadenylase within the CCR4-NOT complex, results in mRNA decay defects during metaphase I (MI) entry. Knockout of B-cell translocation gene-4 (Btg4), an adaptor protein of the CCR4-NOT complex, results in mRNA decay defects following fertilization. Therefore, mechanisms controlling mRNA turnover have significant impacts on oocyte competence and early embryonic development. Post-transcriptional inosine RNA modifications can impact mRNA stability, possibly through a translation mechanism. Here, we assessed inosine RNA modifications in oocytes, eggs, and embryos from Cnot6l-/- and Btg4-/- mice, which display stabilization of mRNA and over-translation of the stabilized transcripts. If inosine modifications have a role in modulating RNA stability, we hypothesize that in these mutant backgrounds, we would observe changes or a disruption in inosine mRNA modifications. To test this, we used a computational approach to identify inosine RNA modifications in total and polysomal RNA-seq data during meiotic maturation (GV, MI, and MII stages). We observed pronounced depletion of inosine mRNA modifications in samples from Cnot6l-/-, but not in Btg4-/- mice. Additionally, analysis of ribosome-associated RNA revealed clearance of inosine modified mRNA. These observations suggest a novel mechanism of mRNA clearance during oocyte maturation, in which inosine-containing transcripts decay in an independent, but parallel mechanism to CCR4-NOT deadenylation.
登录
查看更多内容
影响因子：
3.7
作者：
Doidge R;Mittal S;Aslam A;Winkler GS
通讯作者：
Winkler GS
影响因子：
2.7
作者：
BACHVAROVA, R;DELEON, V
通讯作者：
DELEON, V
影响因子：
56.9
作者：
BRUSA, R;ZIMMERMANN, F;SPRENGEL, R
通讯作者：
SPRENGEL, R
影响因子：
46.9
作者：
Bray, Nicolas L.;Pimentel, Harold;Pachter, Lior
通讯作者：
Pachter, Lior
DOI：
10.1126/science.aay6912
发表时间：
2020-04-17
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Buschauer R;Matsuo Y;Sugiyama T;Chen YH;Alhusaini N;Sweet T;Ikeuchi K;Cheng J;Matsuki Y;Nobuta R;Gilmozzi A;Berninghausen O;Tesina P;Becker T;Coller J;Inada T;Beckmann R
通讯作者：
Beckmann R