Unknown,Transcriptomics,Genomics,Proteomics

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NSun2-mediated cytosine-5 methylation in Vault non-coding RNA determines its processing into small RNAs [iCLIP]

ABSTRACT: Autosomal-recessive loss of the NSUN2 gene has been recently identified as a causative link to intellectual disability disorders in humans. NSun2 is an RNA methyltransferase modifying cytosine-5 in transfer RNAs (tRNA). Whether NSun2 methylates additional RNA species is currently debated. Here, we adapted the individual-nucleotide resolution UV cross-linking and immunoprecipitation method (iCLIP) to identify NSun2-mediated methylation in RNA transcriptome. We confirm site-specific methylation in tRNA and identify messenger and non-coding RNAs as potential methylation targets for NSun2. Using RNA bisulfite sequencing we establish Vault non-coding RNAs as novel substrates for NSun2 and identified six cytosine-5 methylated sites. Furthermore, we show that loss of cytosine-5 methylation in Vault RNAs causes aberrant processing into argonaute-associating small RNA fragments (svRNA). Thus, impaired Vault non-coding RNA processing may be an important contributor to the etiology of NSUN2-deficieny human disorders. Identification of Nsun2 targets by miCLIP in Embryonic kidney (HEK293) cells

ORGANISM(S): Homo sapiens

SUBMITTER: Shobbir Hussain 

PROVIDER: E-GEOD-44385 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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NSun2-mediated cytosine-5 methylation of vault noncoding RNA determines its processing into regulatory small RNAs.

Hussain Shobbir S   Sajini Abdulrahim A AA   Blanco Sandra S   Dietmann Sabine S   Lombard Patrick P   Sugimoto Yoichiro Y   Paramor Maike M   Gleeson Joseph G JG   Odom Duncan T DT   Ule Jernej J   Frye Michaela M  

Cell reports 20130718 2

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   ...[more]

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