NSun2-Mediated Cytosine-5 Methylation of Vault Noncoding RNA Determines Its Processing into Regulatory Small RNAs
| Autor: | Abdulrahim A. Sajini, Duncan T. Odom, Sandra Blanco, Sabine Dietmann, Shobbir Hussain, Yoichiro Sugimoto, Maike Paramor, Patrick Lombard, Michaela Frye, Joseph G. Gleeson, Jernej Ule |
|---|---|
| Rok vydání: | 2013 |
| Předmět: |
Small RNA
RNA Untranslated RNA methylation Molecular Sequence Data Biology Transfection General Biochemistry Genetics and Molecular Biology 03 medical and health sciences Cytosine Report Chlorocebus aethiops Animals Humans Small nucleolar RNA lcsh:QH301-705.5 RNA-Directed DNA Methylation 030304 developmental biology Genetics 0303 health sciences Base Sequence 030302 biochemistry & molecular biology RNA Methyltransferases DNA Methylation Non-coding RNA Long non-coding RNA 3. Good health RNA silencing HEK293 Cells lcsh:Biology (General) Cardiovascular and Metabolic Diseases COS Cells Transcriptome |
| Zdroj: | Cell Reports Cell Reports, Vol 4, Iss 2, Pp 255-261 (2013) |
| ISSN: | 2211-1247 |
| DOI: | 10.1016/j.celrep.2013.06.029 |
| Popis: | Summary 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. Graphical Abstract Highlights • 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. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|