Cell and Developmental Biology, University of Illinois, Urbana, Illinois, United States of America.
Integrative Biology, University of Illinois, Urbana, Illinois, United States of America.
PLoS One. 2021 Nov 30;16(11):e0260005. doi: 10.1371/journal.pone.0260005. eCollection 2021.
Fragile X syndrome results from the loss of expression of the Fragile X Mental Retardation Protein (FMRP). FMRP and RNA helicase Moloney Leukemia virus 10 (MOV10) are important Argonaute (AGO) cofactors for miRNA-mediated translation regulation. We previously showed that MOV10 functionally associates with FMRP. Here we quantify the effect of reduced MOV10 and FMRP expression on dendritic morphology. Murine neurons with reduced MOV10 and FMRP phenocopied Dicer1 KO neurons which exhibit impaired dendritic maturation Hong J (2013), leading us to hypothesize that MOV10 and FMRP regulate DICER expression. In cells and tissues expressing reduced MOV10 or no FMRP, DICER expression was significantly reduced. Moreover, the Dicer1 mRNA is a Cross-Linking Immunoprecipitation (CLIP) target of FMRP Darnell JC (2011), MOV10 Skariah G (2017) and AGO2 Kenny PJ (2020). MOV10 and FMRP modulate expression of DICER1 mRNA through its 3'untranslated region (UTR) and introduction of a DICER1 transgene restores normal neurite outgrowth in the Mov10 KO neuroblastoma Neuro2A cell line and branching in MOV10 heterozygote neurons. Moreover, we observe a global reduction in AGO2-associated microRNAs isolated from Fmr1 KO brain. We conclude that the MOV10-FMRP-AGO2 complex regulates DICER expression, revealing a novel mechanism for regulation of miRNA production required for normal neuronal morphology.
脆性 X 综合征是由于脆性 X 智力迟钝蛋白 (FMRP) 表达缺失引起的。FMRP 和 RNA 解旋酶莫洛尼白血病病毒 10 (MOV10) 是 miRNA 介导的翻译调控的重要 Argonaute (AGO) 辅助因子。我们之前表明,MOV10 与 FMRP 具有功能相关性。在这里,我们量化了降低 MOV10 和 FMRP 表达对树突形态的影响。减少 MOV10 和 FMRP 表达的小鼠神经元与 Dicer1 KO 神经元表型相似,后者表现出树突成熟受损 Hong J (2013),这使我们假设 MOV10 和 FMRP 调节 DICER 的表达。在表达减少的 MOV10 或没有 FMRP 的细胞和组织中,DICER 的表达显著降低。此外,Dicer1 mRNA 是 FMRP Darnell JC (2011)、MOV10 Skariah G (2017) 和 AGO2 Kenny PJ (2020) 的交联免疫沉淀 (CLIP) 靶标。MOV10 和 FMRP 通过 DICER1 mRNA 的 3'非翻译区 (UTR) 调节 DICER1 mRNA 的表达,并且引入 DICER1 转基因可恢复 Mov10 KO 神经母细胞瘤 Neuro2A 细胞系中的正常神经突生长和 MOV10 杂合神经元中的分支。此外,我们观察到从 Fmr1 KO 大脑中分离的 AGO2 相关 microRNAs 整体减少。我们得出结论,MOV10-FMRP-AGO2 复合物调节 DICER 的表达,揭示了 miRNA 产生的新调节机制,这对于正常神经元形态是必需的。