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The modified RNA base acp3U is an attachment site for N-glycans in glycoRNA

Authors
Yixuan Xie,Helena Hemberger
Nicholas A. Till,Peiyuan Chai,Christopher Watkins,Charlotta G. Lebedenko,Reese M. Caldwell,Benson George,Carolyn R. Bertozzi,Benjamin A. Garcia,Ryan A Flynn,Nicholas Till,Charlotta Lebedenko,Reese Caldwell,Carolyn Bertozzi,Benjamin Garcia
+14 authors
,Ryan Flynn
Published
Jan 1, 2023
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Abstract

We recently identified glycoRNA-a previously undescribed glycoconjugate-which consists of RNAs modified with secretory N-glycans and presented on the cell surface. While previous work supported a covalent linkage between RNA and glycans, the direct chemical nature of the RNA-glycan connection was not described. Here we develop a sensitive and scalable protocol to detect and characterize native glycoRNAs. Leveraging periodate oxidation and aldehyde ligation (rPAL) and Sequential Window Acquisition of all Theoretical Mass Spectra (SWATH-MS), we identified the modified RNA base 3-(3-amino-3-carboxypropyl)uridine (acp3U) as a site of attachment of N-glycans in glycoRNA. The sensitivity and robustness of rPAL provided the first evidence of a direct glycan-RNA linkage, and its flexibility will enable further characterization of glycoRNA biology.

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