Direct RNA sequencing reveals m6A modifications on adenovirus RNA are necessary for efficient splicing.

Journal Article (Journal Article)

Adenovirus is a nuclear replicating DNA virus reliant on host RNA processing machinery. Processing and metabolism of cellular RNAs can be regulated by METTL3, which catalyzes the addition of N6-methyladenosine (m6A) to mRNAs. While m6A-modified adenoviral RNAs have been previously detected, the location and function of this mark within the infectious cycle is unknown. Since the complex adenovirus transcriptome includes overlapping spliced units that would impede accurate m6A mapping using short-read sequencing, here we profile m6A within the adenovirus transcriptome using a combination of meRIP-seq and direct RNA long-read sequencing to yield both nucleotide and transcript-resolved m6A detection. Although both early and late viral transcripts contain m6A, depletion of m6A writer METTL3 specifically impacts viral late transcripts by reducing their splicing efficiency. These data showcase a new technique for m6A discovery within individual transcripts at nucleotide resolution, and highlight the role of m6A in regulating splicing of a viral pathogen.

Full Text

Duke Authors

Cited Authors

  • Price, AM; Hayer, KE; McIntyre, ABR; Gokhale, NS; Abebe, JS; Della Fera, AN; Mason, CE; Horner, SM; Wilson, AC; Depledge, DP; Weitzman, MD

Published Date

  • November 26, 2020

Published In

Volume / Issue

  • 11 / 1

Start / End Page

  • 6016 -

PubMed ID

  • 33243990

Pubmed Central ID

  • PMC7691994

Electronic International Standard Serial Number (EISSN)

  • 2041-1723

Digital Object Identifier (DOI)

  • 10.1038/s41467-020-19787-6

Language

  • eng

Conference Location

  • England