Wang Xiaohui, Chang Liang, Wang Huanru, Su Airong, Wu Zhiwei
1 Department of Hematology, Affiliated Nanjing Drum Tower Hospital, Medical School, Nanjing University , Nanjing, People's Republic of China .
2 Center for Public Health Research, Medical School, Nanjing University , Nanjing, People's Republic of China .
DNA Cell Biol. 2017 Jul;36(7):565-570. doi: 10.1089/dna.2017.3633. Epub 2017 May 10.
P-bodies are cytoplasmic foci composed of mRNAs and enzymes involved in mRNA degradation. P-bodies have been found to link to RNA interference and RNA decay mediated by microRNAs (miRNAs) and translational repression. Here, we aim to investigate different effects of overexpressed Dcp1a or GW182 on cytoplasmic aggregates formation and influence on miRNA pathway. Small RNAs were recruited into endogenous foci of P-bodies and aggregates formed by Dcpa1 and GW182 overexpression. However, only overexpressed Dcp1a but not GW182 was colocalized with DDX6, another component of P-bodies and suppressed protein translation. In addition, we investigated the relationship between stress granules and miRNA pathway and found that granules induced by G3BP1 overexpression could recruit small RNAs into the granules and repressed protein translation. As Ago2 is a key component of RNA-induced silencing complex, we also investigated the localization of endogenous Ago2 (edo-Ago2) after Dcp1a and GW182 overexpression, and found that endo-Ago2 did not colocalize with the aggregates induced by overexpression of Dcpla, GW182, and G3BP1. Notably, the ability of miRNA to regulate its target was enhanced by the granules induced by Dcp1a and G3BP1 expression. Our results suggest that overexpressed Dcp1a and GW182 can form different cytoplasmic aggregates and play distinct biological roles in the miRNA pathway.
P小体是由参与mRNA降解的mRNA和酶组成的细胞质聚集体。已发现P小体与RNA干扰以及由微小RNA(miRNA)介导的RNA衰变和翻译抑制有关。在此,我们旨在研究过表达的Dcp1a或GW182对细胞质聚集体形成的不同影响以及对miRNA途径的影响。小RNA被招募到P小体的内源性聚集体以及由Dcpa1和GW182过表达形成的聚集体中。然而,只有过表达的Dcp1a而非GW182与P小体的另一个组分DDX6共定位,并抑制蛋白质翻译。此外,我们研究了应激颗粒与miRNA途径之间的关系,发现由G3BP1过表达诱导的颗粒可将小RNA招募到颗粒中并抑制蛋白质翻译。由于Ago2是RNA诱导沉默复合体的关键组分,我们还研究了Dcp1a和GW182过表达后内源性Ago2(endo-Ago2)的定位,发现endo-Ago2与由Dcpla、GW182和G3BP1过表达诱导的聚集体不共定位。值得注意的是,由Dcp1a和G3BP1表达诱导的颗粒增强了miRNA调节其靶标的能力。我们的结果表明,过表达的Dcp1a和GW182可形成不同的细胞质聚集体,并在miRNA途径中发挥不同的生物学作用。