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Title:
Direct RNA sequencing reveals m6A modifications on adenovirus RNA are necessary for efficient splicing | Nature Communications
Description:
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. Adenovirus transcripts contain N6-methyladenosine (m6A) modification. Here the authors profile m6A modification sites on adenovirus mRNAs using Illumina meRIP-Seq and nanopore direct RNA sequencing, and showcase a role for m6A in splicing of viral late mRNAs.
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headline:Direct RNA sequencing reveals m6A modifications on adenovirus RNA are necessary for efficient splicing
description: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. Adenovirus transcripts contain N6-methyladenosine (m6A) modification. Here the authors profile m6A modification sites on adenovirus mRNAs using Illumina meRIP-Seq and nanopore direct RNA sequencing, and showcase a role for m6A in splicing of viral late mRNAs.
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name:The Childrenâs Hospital of Philadelphia
address:
name:Division of Protective Immunity and Department of Pathology and Laboratory Medicine, The Childrenâs Hospital of Philadelphia, Philadelphia, USA
type:PostalAddress
type:Organization
name:University of Pennsylvania Perelman School of Medicine
address:
name:Department of Pathology and Laboratory Medicine, University of Pennsylvania Perelman School of Medicine, Philadelphia, USA
type:PostalAddress
type:Organization
email:[email protected]
PostalAddress:
name:Division of Protective Immunity and Department of Pathology and Laboratory Medicine, The Childrenâs Hospital of Philadelphia, Philadelphia, USA
name:Department of Biomedical and Health Informatics, The Childrenâs Hospital of Philadelphia, Philadelphia, USA
name:Department of Physiology and Biophysics, Weill Cornell Medicine, New York, USA
name:Tri-Institutional Program in Computational Biology and Medicine, New York, USA
name:Department of Molecular Life Sciences, University of Zurich, Zurich, Switzerland
name:Department of Molecular Genetics and Microbiology, Duke University Medical Center, Durham, USA
name:Department of Immunology, University of Washington, Seattle, USA
name:Department of Medicine, New York University School of Medicine, New York, USA
name:Division of Protective Immunity and Department of Pathology and Laboratory Medicine, The Childrenâs Hospital of Philadelphia, Philadelphia, USA
name:Biological Sciences Graduate Group, University of Maryland, College Park, USA
name:Department of Physiology and Biophysics, Weill Cornell Medicine, New York, USA
name:The HRH Prince Alwaleed Bin Talal Abdulaziz Alsaud Institute for Computational Biomedicine, Weill Cornell Medicine, New York, USA
name:The World Quant Initiative for Quantitative Prediction, Weill Cornell Medicine, New York, USA
name:The Feil Family Brain and Mind Research Institute, Weill Cornell Medicine, New York, USA
name:Department of Molecular Genetics and Microbiology, Duke University Medical Center, Durham, USA
name:Department of Medicine, Duke University Medical Center, Durham, USA
name:Department of Microbiology, New York University School of Medicine, New York, USA
name:Department of Medicine, New York University School of Medicine, New York, USA
name:Division of Protective Immunity and Department of Pathology and Laboratory Medicine, The Childrenâs Hospital of Philadelphia, Philadelphia, USA
name:Department of Pathology and Laboratory Medicine, University of Pennsylvania Perelman School of Medicine, Philadelphia, USA
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