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| Item Type: | Article |
|---|---|
| Title: | NSun2-mediated cytosine-5 methylation of vault noncoding RNA determines its processing into regulatory small RNAs |
| Creators Name: | Hussain, S., Sajini, A.A., Blanco, S., Dietmann, S., Lombard, P., Sugimoto, Y., Paramor, M., Gleeson, J.G., Odom, D.T., Ule, J. and Frye, M. |
| Abstract: | 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. |
| Keywords: | Base Sequence, COS Cell, Cytosine, DNA Methylation, HEK293 Cells, Methyltransferases, Molecular Sequence Data, RNA, Transcriptome, Transfection, Untranslated RNA, Animals, Chlorocebus aethiops |
| Source: | Cell Reports |
| ISSN: | 2211-1247 |
| Publisher: | Cell Press / Elsevier |
| Volume: | 4 |
| Number: | 2 |
| Page Range: | 255-261 |
| Date: | 25 July 2013 |
| Official Publication: | https://doi.org/10.1016/j.celrep.2013.06.029 |
| PubMed: | View item in PubMed |
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