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Attenuation of IFN signaling due to m(6)A modification of the host epitranscriptome promotes EBV lytic reactivation

BACKGROUND: Reactivation of Epstein Barr virus (EBV) leads to modulation of the viral and cellular epitranscriptome. N6-methyladenosine (m(6)A) modification is a type of RNA modification that regulates metabolism of mRNAs. Previous reports demonstrated that m(6)A modification affects the stability a...

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Autores principales: Bose, Dipayan, Lin, Xiang, Gao, Le, Wei, Zhi, Pei, Yonggang, Robertson, Erle S.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10012557/
https://www.ncbi.nlm.nih.gov/pubmed/36918845
http://dx.doi.org/10.1186/s12929-023-00911-9
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author Bose, Dipayan
Lin, Xiang
Gao, Le
Wei, Zhi
Pei, Yonggang
Robertson, Erle S.
author_facet Bose, Dipayan
Lin, Xiang
Gao, Le
Wei, Zhi
Pei, Yonggang
Robertson, Erle S.
author_sort Bose, Dipayan
collection PubMed
description BACKGROUND: Reactivation of Epstein Barr virus (EBV) leads to modulation of the viral and cellular epitranscriptome. N6-methyladenosine (m(6)A) modification is a type of RNA modification that regulates metabolism of mRNAs. Previous reports demonstrated that m(6)A modification affects the stability and metabolism of EBV encoded mRNAs. However, the effect of reactivation on reprograming of the cellular mRNAs, and how this contributes to successful induction of lytic reactivation is not known. METHODS: Methylated RNA immunoprecipitation sequencing (MeRIP-seq), transcriptomic RNA sequencing (RNA-seq) and RNA pull-down PCR were used to screen and validate differentially methylated targets. Western blotting, quantitative real-time PCR (RT-qPCR) and immunocytochemistry were used to investigate the expression and localization of different proteins. RNA stability and polysome analysis assays were used to detect the half-lives and translation efficiencies of downstream genes. Insertion of point mutation to disrupt the m(6)A methylation sites was used to verify the effect of m(6)A methylation on its stability and expression levels. RESULTS: We report that during EBV reactivation the m(6)A eraser ALKBH5 is significantly downregulated leading to enhanced methylation of the cellular transcripts DTX4 and TYK2, that results in degradation of TYK2 mRNAs and higher efficiency of translation of DTX4 mRNAs. This resulted in attenuation of IFN signaling that promoted progression of viral lytic replication. Furthermore, inhibition of m(6)A methylation of these transcripts led to increased production of IFN, and a substantial reduction in viral copy number, which suggests abrogation of lytic viral replication. CONCLUSION: Our findings illuminate the significance of m(6)A modification in overcoming the innate immune response during EBV reactivation. We now report that during lytic reactivation EBV targets the RNA methylation system of the host to attenuate the innate immune response by suppressing the interferon signaling which facilitates successful lytic replication of the virus. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12929-023-00911-9.
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spelling pubmed-100125572023-03-15 Attenuation of IFN signaling due to m(6)A modification of the host epitranscriptome promotes EBV lytic reactivation Bose, Dipayan Lin, Xiang Gao, Le Wei, Zhi Pei, Yonggang Robertson, Erle S. J Biomed Sci Research BACKGROUND: Reactivation of Epstein Barr virus (EBV) leads to modulation of the viral and cellular epitranscriptome. N6-methyladenosine (m(6)A) modification is a type of RNA modification that regulates metabolism of mRNAs. Previous reports demonstrated that m(6)A modification affects the stability and metabolism of EBV encoded mRNAs. However, the effect of reactivation on reprograming of the cellular mRNAs, and how this contributes to successful induction of lytic reactivation is not known. METHODS: Methylated RNA immunoprecipitation sequencing (MeRIP-seq), transcriptomic RNA sequencing (RNA-seq) and RNA pull-down PCR were used to screen and validate differentially methylated targets. Western blotting, quantitative real-time PCR (RT-qPCR) and immunocytochemistry were used to investigate the expression and localization of different proteins. RNA stability and polysome analysis assays were used to detect the half-lives and translation efficiencies of downstream genes. Insertion of point mutation to disrupt the m(6)A methylation sites was used to verify the effect of m(6)A methylation on its stability and expression levels. RESULTS: We report that during EBV reactivation the m(6)A eraser ALKBH5 is significantly downregulated leading to enhanced methylation of the cellular transcripts DTX4 and TYK2, that results in degradation of TYK2 mRNAs and higher efficiency of translation of DTX4 mRNAs. This resulted in attenuation of IFN signaling that promoted progression of viral lytic replication. Furthermore, inhibition of m(6)A methylation of these transcripts led to increased production of IFN, and a substantial reduction in viral copy number, which suggests abrogation of lytic viral replication. CONCLUSION: Our findings illuminate the significance of m(6)A modification in overcoming the innate immune response during EBV reactivation. We now report that during lytic reactivation EBV targets the RNA methylation system of the host to attenuate the innate immune response by suppressing the interferon signaling which facilitates successful lytic replication of the virus. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12929-023-00911-9. BioMed Central 2023-03-14 /pmc/articles/PMC10012557/ /pubmed/36918845 http://dx.doi.org/10.1186/s12929-023-00911-9 Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research
Bose, Dipayan
Lin, Xiang
Gao, Le
Wei, Zhi
Pei, Yonggang
Robertson, Erle S.
Attenuation of IFN signaling due to m(6)A modification of the host epitranscriptome promotes EBV lytic reactivation
title Attenuation of IFN signaling due to m(6)A modification of the host epitranscriptome promotes EBV lytic reactivation
title_full Attenuation of IFN signaling due to m(6)A modification of the host epitranscriptome promotes EBV lytic reactivation
title_fullStr Attenuation of IFN signaling due to m(6)A modification of the host epitranscriptome promotes EBV lytic reactivation
title_full_unstemmed Attenuation of IFN signaling due to m(6)A modification of the host epitranscriptome promotes EBV lytic reactivation
title_short Attenuation of IFN signaling due to m(6)A modification of the host epitranscriptome promotes EBV lytic reactivation
title_sort attenuation of ifn signaling due to m(6)a modification of the host epitranscriptome promotes ebv lytic reactivation
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10012557/
https://www.ncbi.nlm.nih.gov/pubmed/36918845
http://dx.doi.org/10.1186/s12929-023-00911-9
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