NSun2-mediated cytosine-5 methylation of vault noncoding RNA determines its processing into regulatory small RNAs.
NSun2-mediated cytosine-5 methylation of vault noncoding RNA determines its processing into regulatory small RNAs.
复制标题
DOI：
10.1016/j.celrep.2013.06.029
复制
发表时间：
2013-07-25
期刊：
影响因子：
8.8
通讯作者：
Frye M
中科院分区：
文献类型：
--
作者：
Hussain S;Sajini AA;Blanco S;Dietmann S;Lombard P;Sugimoto Y;Paramor M;Gleeson JG;Odom DT;Ule J;Frye M
NSUN2基因的常染色体不必要的损失已被确定为与人体中智障疾病的关键联系。由于缺乏敏感和可靠的分子技术，我们将MICLIP描述为识别RNA甲基化位点的另一种方法转录组是单个核苷酸 - 分辨率交联和免疫沉淀（ICLIP）的定制版本。胞质，其中三种是通过缺乏NSUN2蛋白的患者细胞来确认的。表明，在拱顶RNA中的胞嘧啶-5甲基化的损失会导致异常处理与Argonaute相关的小RNA片段，可以用作microRNA，而保管NCRNA的处理受损可能有助于NSUN2缺乏性的人类疾病的病因。
MICLIP检测RNA中NSUN2介导的胞嘧啶-5甲基化
金库非编码RNA被NSUN2甲基化
cytosine-5中的RNA中确定其处理成小RNA（SVRNA）
SVRNA与Argonaute蛋白结合并暴露于MicroRNA样函数
在此工作中缺乏敏感和可靠的分子技术，对RNA转录组中的胞嘧啶5甲基化的全面分析受到了阻碍。 RNA甲基转移酶的位置，NSUN2进一步展示了NSUN2介导的金库NCRNA的甲基化如何影响其加工成小的microRNA样分子。
Autosomal-recessive loss of the NSUN2 gene has been identified as a causative link to intellectual disability disorders in humans. NSun2 is an RNA methyltransferase modifying cytosine-5 in transfer RNAs (tRNAs), yet the identification of cytosine methylation in other RNA species has been hampered by the lack of sensitive and reliable molecular techniques. Here, we describe miCLIP as an additional approach for identifying RNA methylation sites in transcriptomes. miCLIP is a customized version of the individual-nucleotide-resolution crosslinking and immunoprecipitation (iCLIP) method. We confirm site-specific methylation in tRNAs and additional messenger and noncoding RNAs (ncRNAs). Among these, vault ncRNAs contained six NSun2-methylated cytosines, three of which were confirmed by RNA bisulfite sequencing. Using patient cells lacking the NSun2 protein, we further show that loss of cytosine-5 methylation in vault RNAs causes aberrant processing into Argonaute-associated small RNA fragments that can function as microRNAs. Thus, impaired processing of vault ncRNA may contribute to the etiology of NSun2-deficiency human disorders. miCLIP detects NSun2-mediated cytosine-5 methylation in RNA Vault noncoding RNA is methylated by NSun2 Cytosine-5 in Vault RNA determines its processing into small RNA (svRNA) svRNAs bind to Argonaute proteins and exhibit microRNA-like functions Comprehensive analyses of cytosine-5 methylation in the RNA transcriptome have previously been hampered by the lack of sensitive and reliable molecular techniques. In this work, Ule, Frye, and colleagues describe the methylation-iCLIP (miCLIP) method that they used to identify target sites of the RNA methyltransferase, NSun2. Among the targeted noncoding RNAs were vault RNAs, previously associated with resistance to chemotherapy in cancer. They further show how NSun2-mediated methylation of vault ncRNAs influences their processing into small microRNA-like molecules.
登录
查看更多内容
影响因子：
14.9
作者：
Skreka K;Schafferer S;Nat IR;Zywicki M;Salti A;Apostolova G;Griehl M;Rederstorff M;Dechant G;Hüttenhofer A
通讯作者：
Hüttenhofer A
影响因子：
10.7
作者：
Stadler, Peter F.;Chen, Julian J. -L.;Ullmann, Kerstin
通讯作者：
Ullmann, Kerstin
影响因子：
10.5
作者：
Schaefer, Matthias;Pollex, Tim;Lyko, Frank
通讯作者：
Lyko, Frank
影响因子：
56.9
作者：
Goll, MG;Kirpekar, F;Bestor, TH
通讯作者：
Bestor, TH
影响因子：
16.8
作者：
通讯作者：
--