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

Publication ,  Journal Article
Price, AM; Hayer, KE; McIntyre, ABR; Gokhale, NS; Abebe, JS; Della Fera, AN; Mason, CE; Horner, SM; Wilson, AC; Depledge, DP; Weitzman, MD
Published in: Nat Commun
November 26, 2020

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.

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Published In

Nat Commun

DOI

EISSN

2041-1723

Publication Date

November 26, 2020

Volume

11

Issue

1

Start / End Page

6016

Location

England

Related Subject Headings

  • Virus Replication
  • Sequence Analysis, RNA
  • RNA, Viral
  • RNA, Small Interfering
  • RNA Splicing
  • Methyltransferases
  • Humans
  • Host-Pathogen Interactions
  • HEK293 Cells
  • Gene Knockout Techniques
 

Citation

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Price, A. M., Hayer, K. E., McIntyre, A. B. R., Gokhale, N. S., Abebe, J. S., Della Fera, A. N., … Weitzman, M. D. (2020). Direct RNA sequencing reveals m6A modifications on adenovirus RNA are necessary for efficient splicing. Nat Commun, 11(1), 6016. https://doi.org/10.1038/s41467-020-19787-6
Price, Alexander M., Katharina E. Hayer, Alexa B. R. McIntyre, Nandan S. Gokhale, Jonathan S. Abebe, Ashley N. Della Fera, Christopher E. Mason, et al. “Direct RNA sequencing reveals m6A modifications on adenovirus RNA are necessary for efficient splicing.Nat Commun 11, no. 1 (November 26, 2020): 6016. https://doi.org/10.1038/s41467-020-19787-6.
Price AM, Hayer KE, McIntyre ABR, Gokhale NS, Abebe JS, Della Fera AN, et al. Direct RNA sequencing reveals m6A modifications on adenovirus RNA are necessary for efficient splicing. Nat Commun. 2020 Nov 26;11(1):6016.
Price, Alexander M., et al. “Direct RNA sequencing reveals m6A modifications on adenovirus RNA are necessary for efficient splicing.Nat Commun, vol. 11, no. 1, Nov. 2020, p. 6016. Pubmed, doi:10.1038/s41467-020-19787-6.
Price AM, Hayer KE, McIntyre ABR, Gokhale NS, Abebe JS, Della Fera AN, Mason CE, Horner SM, Wilson AC, Depledge DP, Weitzman MD. Direct RNA sequencing reveals m6A modifications on adenovirus RNA are necessary for efficient splicing. Nat Commun. 2020 Nov 26;11(1):6016.

Published In

Nat Commun

DOI

EISSN

2041-1723

Publication Date

November 26, 2020

Volume

11

Issue

1

Start / End Page

6016

Location

England

Related Subject Headings

  • Virus Replication
  • Sequence Analysis, RNA
  • RNA, Viral
  • RNA, Small Interfering
  • RNA Splicing
  • Methyltransferases
  • Humans
  • Host-Pathogen Interactions
  • HEK293 Cells
  • Gene Knockout Techniques