Whitehead Institute for Biomedical Research, Cambridge, MA, 02142, USA.
Howard Hughes Medical Institute, Cambridge, MA, 02142, USA.
Genome Biol. 2017 Oct 31;18(1):211. doi: 10.1186/s13059-017-1330-z.
All mRNAs are bound in vivo by proteins to form mRNA-protein complexes (mRNPs), but changes in the composition of mRNPs during posttranscriptional regulation remain largely unexplored. Here, we have analyzed, on a transcriptome-wide scale, how microRNA-mediated repression modulates the associations of the core mRNP components eIF4E, eIF4G, and PABP and of the decay factor DDX6 in human cells.
Despite the transient nature of repressed intermediates, we detect significant changes in mRNP composition, marked by dissociation of eIF4G and PABP, and by recruitment of DDX6. Furthermore, although poly(A)-tail length has been considered critical in post-transcriptional regulation, differences in steady-state tail length explain little of the variation in either PABP association or mRNP organization more generally. Instead, relative occupancy of core components correlates best with gene expression.
These results indicate that posttranscriptional regulatory factors, such as microRNAs, influence the associations of PABP and other core factors, and do so without substantially affecting steady-state tail length.
所有的 mRNA 在体内都与蛋白质结合形成 mRNA-蛋白质复合物(mRNP),但在转录后调控过程中 mRNP 组成的变化在很大程度上仍未得到探索。在这里,我们在转录组范围内分析了 microRNA 介导的抑制如何调节核心 mRNP 成分 eIF4E、eIF4G 和 PABP 以及降解因子 DDX6 在人类细胞中的关联。
尽管抑制中间产物具有瞬时性质,但我们检测到 mRNP 组成的显著变化,其特征是 eIF4G 和 PABP 的解离,以及 DDX6 的募集。此外,尽管 poly(A)-尾长在转录后调控中被认为是至关重要的,但在 PABP 结合或更普遍的 mRNP 组织方面,稳态尾长的差异解释不了多少变化。相反,核心成分的相对占有率与基因表达相关性最好。
这些结果表明,转录后调控因子,如 microRNAs,影响 PABP 和其他核心因子的关联,而且在不显著影响稳态尾长的情况下做到这一点。