Rosalind and Morris Goodman Cancer Research Centre, McGill University, Montreal H3A 1A3 QC, Canada.
Mol Cell. 2010 Nov 24;40(4):558-70. doi: 10.1016/j.molcel.2010.11.003.
To understand how miRNA-mediated silencing impacts on embryonic mRNAs, we conducted a functional survey of abundant maternal and zygotic miRNA families in the C. elegans embryo. We show that the miR-35-42 and the miR-51-56 miRNA families define maternal and zygotic miRNA-induced silencing complexes (miRISCs), respectively, that share a large number of components. Using a cell-free C. elegans embryonic extract, we demonstrate that the miRISC directs the rapid deadenylation of reporter mRNAs with natural 3'UTRs. The deadenylated targets are translationally suppressed and remarkably stable. Sampling of the predicted miR-35-42 targets reveals that roughly half are deadenylated in a miRNA-dependent manner, but with each target displaying a distinct efficiency and pattern of deadenylation. Finally, we demonstrate that functional cooperation between distinct miRISCs within 3'UTRs is required to potentiate deadenylation. With this report, we reveal the extensive and direct impact of miRNA-mediated deadenylation on embryonic mRNAs.
为了了解 miRNA 介导的沉默对胚胎 mRNA 的影响,我们对秀丽隐杆线虫胚胎中丰富的母源和合子 miRNA 家族进行了功能调查。我们表明,miR-35-42 和 miR-51-56 miRNA 家族分别定义了母源和合子 miRNA 诱导的沉默复合物 (miRISCs),它们共享大量的成分。使用无细胞秀丽隐杆线虫胚胎提取物,我们证明了 miRISC 指导报告 mRNA 与天然 3'UTR 快速去腺苷酸化。去腺苷酸化的靶标被翻译抑制,并显著稳定。对预测的 miR-35-42 靶标的采样表明,大约一半以 miRNA 依赖的方式被去腺苷酸化,但每个靶标显示出不同的去腺苷酸化效率和模式。最后,我们证明了不同的 miRISCs 在 3'UTR 内的功能合作对于增强去腺苷酸化是必需的。通过本报告,我们揭示了 miRNA 介导的去腺苷酸化对胚胎 mRNA 的广泛而直接的影响。