Coxsackievirus B3-Induced m<sup>6</sup>A Modification of RNA Enhances Viral Replication via Suppression of YTHDF-Mediated Stress Granule Formation
N6-methyladenosine (m<sup>6</sup>A) is the most prevalent internal RNA modification. Here, we demonstrate that coxsackievirus B3 (CVB3), a common causative agent of viral myocarditis, induces m<sup>6</sup>A modification primarily at the stop codon and 3′ untranslated regions...
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2024-10-01
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| author | Guangze Zhao Huifang M. Zhang Yankuan T. Chen Kerry Shi Sana Aghakeshmiri Fione Yip Honglin Luo Bruce McManus Decheng Yang |
| author_facet | Guangze Zhao Huifang M. Zhang Yankuan T. Chen Kerry Shi Sana Aghakeshmiri Fione Yip Honglin Luo Bruce McManus Decheng Yang |
| author_sort | Guangze Zhao |
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| description | N6-methyladenosine (m<sup>6</sup>A) is the most prevalent internal RNA modification. Here, we demonstrate that coxsackievirus B3 (CVB3), a common causative agent of viral myocarditis, induces m<sup>6</sup>A modification primarily at the stop codon and 3′ untranslated regions of its genome. As a positive-sense single-stranded RNA virus, CVB3 replicates exclusively in the cytoplasm through a cap-independent translation initiation mechanism. Our study shows that CVB3 modulates the expression and nucleo-cytoplasmic transport of the m<sup>6</sup>A machinery components—METTL3, ALKBH5 and YTHDFs—resulting in increased m<sup>6</sup>A modifications that enhance viral replication. Mechanistically, this enhancement is mediated through YTHDF-driven stress granule (SG) formation. We observed that YTHDF proteins co-localize with human antigen R (HuR), a protein facilitating cap-independent translation, in SGs during early infection. Later in infection, YTHDFs are cleaved, suppressing SG formation. Notably, for the first time, we identified that during early infection CVB3’s RNA-dependent RNA polymerase (3D) and double-stranded RNA (dsRNA) are stored in SGs, co-localizing with HuR. This early-stage sequestration likely protects viral components for use in late-phase replication, when SGs are disrupted due to YTHDF cleavage. In summary, our findings reveal that CVB3-induced m<sup>6</sup>A modifications enhance viral replication by regulating YTHDF-mediated SG dynamics. This study provides a potential therapeutic strategy for CVB3-induced myocarditis. |
| format | Article |
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| issn | 2076-2607 |
| language | English |
| publishDate | 2024-10-01 |
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| series | Microorganisms |
| spelling | doaj-art-e6d3155db8ea44279a378fc9130672d42025-08-20T02:05:06ZengMDPI AGMicroorganisms2076-26072024-10-011211215210.3390/microorganisms12112152Coxsackievirus B3-Induced m<sup>6</sup>A Modification of RNA Enhances Viral Replication via Suppression of YTHDF-Mediated Stress Granule FormationGuangze Zhao0Huifang M. Zhang1Yankuan T. Chen2Kerry Shi3Sana Aghakeshmiri4Fione Yip5Honglin Luo6Bruce McManus7Decheng Yang8Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, BC V6T 1Z4, CanadaDepartment of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, BC V6T 1Z4, CanadaCentre for Heart Lung Innovation, University of British Columbia, St. Paul’s Hospital, 1081 Burrard Street, Vancouver, BC V6Z 1Y6, CanadaCentre for Heart Lung Innovation, University of British Columbia, St. Paul’s Hospital, 1081 Burrard Street, Vancouver, BC V6Z 1Y6, CanadaCentre for Heart Lung Innovation, University of British Columbia, St. Paul’s Hospital, 1081 Burrard Street, Vancouver, BC V6Z 1Y6, CanadaCentre for Heart Lung Innovation, University of British Columbia, St. Paul’s Hospital, 1081 Burrard Street, Vancouver, BC V6Z 1Y6, CanadaDepartment of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, BC V6T 1Z4, CanadaDepartment of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, BC V6T 1Z4, CanadaDepartment of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, BC V6T 1Z4, CanadaN6-methyladenosine (m<sup>6</sup>A) is the most prevalent internal RNA modification. Here, we demonstrate that coxsackievirus B3 (CVB3), a common causative agent of viral myocarditis, induces m<sup>6</sup>A modification primarily at the stop codon and 3′ untranslated regions of its genome. As a positive-sense single-stranded RNA virus, CVB3 replicates exclusively in the cytoplasm through a cap-independent translation initiation mechanism. Our study shows that CVB3 modulates the expression and nucleo-cytoplasmic transport of the m<sup>6</sup>A machinery components—METTL3, ALKBH5 and YTHDFs—resulting in increased m<sup>6</sup>A modifications that enhance viral replication. Mechanistically, this enhancement is mediated through YTHDF-driven stress granule (SG) formation. We observed that YTHDF proteins co-localize with human antigen R (HuR), a protein facilitating cap-independent translation, in SGs during early infection. Later in infection, YTHDFs are cleaved, suppressing SG formation. Notably, for the first time, we identified that during early infection CVB3’s RNA-dependent RNA polymerase (3D) and double-stranded RNA (dsRNA) are stored in SGs, co-localizing with HuR. This early-stage sequestration likely protects viral components for use in late-phase replication, when SGs are disrupted due to YTHDF cleavage. In summary, our findings reveal that CVB3-induced m<sup>6</sup>A modifications enhance viral replication by regulating YTHDF-mediated SG dynamics. This study provides a potential therapeutic strategy for CVB3-induced myocarditis.https://www.mdpi.com/2076-2607/12/11/2152coxsackievirus B3N6-methyladenosineHuR3Dreader protein YTHDFsstress granule |
| spellingShingle | Guangze Zhao Huifang M. Zhang Yankuan T. Chen Kerry Shi Sana Aghakeshmiri Fione Yip Honglin Luo Bruce McManus Decheng Yang Coxsackievirus B3-Induced m<sup>6</sup>A Modification of RNA Enhances Viral Replication via Suppression of YTHDF-Mediated Stress Granule Formation Microorganisms coxsackievirus B3 N6-methyladenosine HuR 3D reader protein YTHDFs stress granule |
| title | Coxsackievirus B3-Induced m<sup>6</sup>A Modification of RNA Enhances Viral Replication via Suppression of YTHDF-Mediated Stress Granule Formation |
| title_full | Coxsackievirus B3-Induced m<sup>6</sup>A Modification of RNA Enhances Viral Replication via Suppression of YTHDF-Mediated Stress Granule Formation |
| title_fullStr | Coxsackievirus B3-Induced m<sup>6</sup>A Modification of RNA Enhances Viral Replication via Suppression of YTHDF-Mediated Stress Granule Formation |
| title_full_unstemmed | Coxsackievirus B3-Induced m<sup>6</sup>A Modification of RNA Enhances Viral Replication via Suppression of YTHDF-Mediated Stress Granule Formation |
| title_short | Coxsackievirus B3-Induced m<sup>6</sup>A Modification of RNA Enhances Viral Replication via Suppression of YTHDF-Mediated Stress Granule Formation |
| title_sort | coxsackievirus b3 induced m sup 6 sup a modification of rna enhances viral replication via suppression of ythdf mediated stress granule formation |
| topic | coxsackievirus B3 N6-methyladenosine HuR 3D reader protein YTHDFs stress granule |
| url | https://www.mdpi.com/2076-2607/12/11/2152 |
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