PT  - JOURNAL ARTICLE
AU  - Alexander M. Price
AU  - Katharina E. Hayer
AU  - Alexa B.R. McIntyre
AU  - Nandan S. Gokhale
AU  - Jonathan S. Abebe
AU  - Ashley N. Della Fera
AU  - Christopher E. Mason
AU  - Stacy M. Horner
AU  - Angus C. Wilson
AU  - Daniel P. Depledge
AU  - Matthew D. Weitzman
TI  - Direct RNA sequencing reveals m<sup>6</sup>A modifications on adenovirus RNA are necessary for efficient splicing
AID  - 10.1101/865485
DP  - 2020 Jan 01
TA  - bioRxiv
PG  - 865485
4099  - http://biorxiv.org/content/early/2020/08/13/865485.short
4100  - http://biorxiv.org/content/early/2020/08/13/865485.full
AB  - 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, we profiled 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.Competing Interest StatementC.E.M. is a cofounder and board member for Biotia and Onegevity Health, as well as an advisor or compensated speaker for Abbvie, Acuamark Diagnostics, ArcBio, BioRad, DNA Genotek, Genialis, Genpro, Karius, Illumina, New England Biolabs, QIAGEN, Whole Biome, and Zymo Research. The other authors declare no competing interests.