FUS modulates the level of ribosomal RNA modifications by regulating a subset of snoRNA expression [RiboMeth-Seq]

FUS is a nucleocytoplasmic protein involved in many processes such as RNA transport, translation, and metabolism as well as genomic maintenance. FUS mutations are found in several neurodegenerative diseases. We find that FUS knockout (KO) and FUS mutation affects the expression of many C/D box snoRNAs, H/ACA box snoRNAs and scaRNAs. snoRNAs notably serve as adaptors for the 2'-O-methylation (2'-O-me) and pseudouridylation of ribosomal RNA (rRNA). We carried out small RNA sequencing and analysed the impact on ribosomal RNA 2'-O-methylation and pseudouridylation using RiboMeth-Seq and HydraPsiSeq in FUS knock-out model cell lines (HEK293T FUS KO cells and SH-SY5Y (neuroblastoma) FUS KO cells as well as SH-SY5Y cells with FUS R495X mutation).

FUS蛋白（FUS）是一类核质穿梭蛋白，参与RNA转运、翻译、RNA代谢及基因组维持等诸多生物学过程。FUS突变可见于多种神经退行性疾病。本研究发现，FUS基因敲除（KO）与FUS突变可影响大量C/D盒核仁小RNA（C/D box snoRNAs）、H/ACA盒核仁小RNA（H/ACA box snoRNAs）以及小卡哈尔体特异性RNA（scaRNAs）的表达。核仁小RNA（snoRNAs）主要作为适配因子介导核糖体RNA（rRNA）的2'-O-甲基化（2'-O-methylation）与假尿苷化修饰。我们借助小RNA测序技术，并利用RiboMeth-Seq与HydraPsiSeq两种方法，分析了FUS基因敲除模型细胞系——包括HEK293T细胞FUS基因敲除株、SH-SY5Y（神经母细胞瘤）细胞FUS基因敲除株，以及携带FUS R495X突变的SH-SY5Y细胞——中核糖体RNA 2'-O-甲基化与假尿苷化水平的变化。