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

• DOI: 10.1038/s41467-020-19787-6
• Publication Date: 2020-11-26T11:08:08Z
• Authors: Alexander M. Price; Katharina E. Hayer; Alexa B. R. McIntyre; Nandan S. Gokhale; Jonathan S. Abebe; Ashley N. Della Fera; Christopher E. Mason; Stacy M. Horner; Angus C. Wilson; Daniel P. Depledge; Matthew D. Weitzman
• Keywords: 0303 health sciences; Adenosine; Sequence Analysis, RNA; Science; Adenoviruses, Human; RNA Splicing; Q; Methyltransferases; Virus Replication; Article; 3. Good health; Adenovirus Infections, Human; Gene Knockout Techniques; 03 medical and health sciences; HEK293 Cells; A549 Cells; Gene Knockdown Techniques; DNA, Viral; Host-Pathogen Interactions; Humans; RNA, Viral; RNA, Small Interfering

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 N6-methyladenosine (m6A) to mRNAs. While m6A-modified adenoviral have been previously detected, location function this mark within infectious cycle unknown. Since complex adenovirus transcriptome includes overlapping spliced units that would impede accurate m6A mapping using short-read sequencing, here we profile combination meRIP-seq direct long-read sequencing yield both nucleotide transcript-resolved detection. Although early late viral transcripts contain m6A, depletion writer METTL3 specifically impacts reducing their splicing efficiency. These data showcase new technique for discovery individual at resolution, highlight role in regulating pathogen.