Publication Date:
2016-04-08
Description:
Pseudouridine (5-ribosyluracil, ) is the only ‘mass-silent’ nucleoside produced by post-transcriptional RNA modification. We describe here a novel mass spectrometry (MS)-based method for direct determination of in RNA. The method assigns a -containing nucleolytic RNA fragment by an accurate measurement of a signature doubly dehydrated nucleoside anion ([C 9 H 7 N 2 O 4 ] 1– , m/z 207.04) produced by collision-induced dissociation MS, and it determines the -containing nucleotide sequence by pseudo-MS 3 , i.e. in-source fragmentation followed by MS 2 . By applying this method, we identified all of the known s in the canonical human spliceosomal snRNAs and, unexpectedly, found two previously unknown s in the U5 and U6 snRNAs. Because the method allows direct determination of in a subpicomole quantity of RNA, it will serve as a useful tool for the structure/function studies of a wide variety of non-coding RNAs.
Keywords:
Nucleic acid modification, RNA characterisation and manipulation
Print ISSN:
0305-1048
Electronic ISSN:
1362-4962
Topics:
Biology
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