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

Yixuan Xie, Peiyuan Chai, Nicholas A. Till, Helena Hemberger, Charlotta G. Lebedenko, Jennifer Porat, Christopher P. Watkins, Reese M. Caldwell, Benson M. George, Jonathan Perr, Carolyn R. Bertozzi, Benjamin A. Garcia, Ryan A. Flynn

Research output: Contribution to journalArticlepeer-review

9 Scopus citations

Abstract

GlycoRNA consists of RNAs modified with secretory N-glycans that are presented on the cell surface. Although 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 RNA-optimized 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. rPAL offers sensitivity and robustness as an approach for characterizing direct glycan-RNA linkages occurring in cells, and its flexibility will enable further exploration of glycoRNA biology.

Original languageEnglish
Pages (from-to)5228-5237.e12
JournalCell
Volume187
Issue number19
DOIs
StatePublished - Sep 19 2024

Keywords

  • RNA modifications
  • acp3U
  • cell surface
  • glycoRNA

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