SARS-CoV-2 Mac1 is required for IFN antagonism and efficient virus replication in mice Article Swipe
Yousef M. Alhammad
,
Srivatsan Parthasarathy
,
Roshan Ghimire
,
Joseph J. O’Connor
,
Catherine M. Kerr
,
Jessica J. Pfannenstiel
,
Debarati Chanda
,
Caden A. Miller
,
Robert L. Unckless
,
Sonia Zúñiga
,
Luis Enjuanes
,
Sunil More
,
Rudragouda Channappanavar
,
Anthony R. Fehr
·
YOU?
·
· 2023
· Open Access
·
· DOI: https://doi.org/10.1101/2023.04.06.535927
YOU?
·
· 2023
· Open Access
·
· DOI: https://doi.org/10.1101/2023.04.06.535927
Several coronavirus (CoV) encoded proteins are being evaluated as targets for antiviral therapies for COVID-19. Included in this set of proteins is the conserved macrodomain, or Mac1, an ADP-ribosylhydrolase and ADP-ribose binding protein. Utilizing point mutant recombinant viruses, Mac1 was shown to be critical for both murine hepatitis virus (MHV) and severe acute respiratory syndrome (SARS)-CoV virulence. However, as a potential drug target, it is imperative to understand how a complete Mac1 deletion impacts the replication and pathogenesis of different CoVs. To this end, we created recombinant bacterial artificial chromosomes (BACs) containing complete Mac1 deletions (ΔMac1) in MHV, MERS-CoV, and SARS-CoV-2. While we were unable to recover infectious virus from MHV or MERS-CoV ΔMac1 BACs, SARS-CoV-2 ΔMac1 was readily recovered from BAC transfection, indicating a stark difference in the requirement for Mac1 between different CoVs. Furthermore, SARS-CoV-2 ΔMac1 replicated at or near wild-type levels in multiple cell lines susceptible to infection. However, in a mouse model of severe infection, ΔMac1 was quickly cleared causing minimal pathology without any morbidity. ΔMac1 SARS-CoV-2 induced increased levels of interferon (IFN) and interferon-stimulated gene (ISG) expression in cell culture and mice, indicating that Mac1 blocks IFN responses which may contribute to its attenuation. ΔMac1 infection also led to a stark reduction in inflammatory monocytes and neutrophils. These results demonstrate that Mac1 only minimally impacts SARS-CoV-2 replication, unlike MHV and MERS-CoV, but is required for SARS-CoV-2 pathogenesis and is a unique antiviral drug target. SIGNIFICANCE All CoVs, including SARS-CoV-2, encode for a conserved macrodomain (Mac1) that counters host ADP-ribosylation. Prior studies with SARS-CoV-1 and MHV found that Mac1 blocks IFN production and promotes CoV pathogenesis, which has prompted the development of SARS-CoV-2 Mac1 inhibitors. However, development of these compounds into antivirals requires that we understand how SARS-CoV-2 lacking Mac1 replicates and causes disease in vitro and in vivo . Here we found that SARS-CoV-2 containing a complete Mac1 deletion replicates normally in cell culture but induces an elevated IFN response, has reduced viral loads in vivo , and does not cause significant disease in mice. These results will provide a roadmap for testing Mac1 inhibitors, help identify Mac1 functions, and open additional avenues for coronavirus therapies.
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Metadata
- Type
- preprint
- Language
- en
- Landing Page
- https://doi.org/10.1101/2023.04.06.535927
- https://www.biorxiv.org/content/biorxiv/early/2023/04/06/2023.04.06.535927.full.pdf
- OA Status
- green
- Cited By
- 11
- References
- 68
- Related Works
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- OpenAlex ID
- https://openalex.org/W4362700644
All OpenAlex metadata
Raw OpenAlex JSON
- OpenAlex ID
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https://openalex.org/W4362700644Canonical identifier for this work in OpenAlex
- DOI
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https://doi.org/10.1101/2023.04.06.535927Digital Object Identifier
- Title
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SARS-CoV-2 Mac1 is required for IFN antagonism and efficient virus replication in miceWork title
- Type
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preprintOpenAlex work type
- Language
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enPrimary language
- Publication year
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2023Year of publication
- Publication date
-
2023-04-06Full publication date if available
- Authors
-
Yousef M. Alhammad, Srivatsan Parthasarathy, Roshan Ghimire, Joseph J. O’Connor, Catherine M. Kerr, Jessica J. Pfannenstiel, Debarati Chanda, Caden A. Miller, Robert L. Unckless, Sonia Zúñiga, Luis Enjuanes, Sunil More, Rudragouda Channappanavar, Anthony R. FehrList of authors in order
- Landing page
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https://doi.org/10.1101/2023.04.06.535927Publisher landing page
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https://www.biorxiv.org/content/biorxiv/early/2023/04/06/2023.04.06.535927.full.pdfDirect link to full text PDF
- Open access
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YesWhether a free full text is available
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greenOpen access status per OpenAlex
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https://www.biorxiv.org/content/biorxiv/early/2023/04/06/2023.04.06.535927.full.pdfDirect OA link when available
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Antagonism, Replication (statistics), Virology, Viral replication, Biology, Virus, Genetics, ReceptorTop concepts (fields/topics) attached by OpenAlex
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11Total citation count in OpenAlex
- Citations by year (recent)
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2025: 1, 2024: 2, 2023: 8Per-year citation counts (last 5 years)
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68Number of works referenced by this work
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10Other works algorithmically related by OpenAlex
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| abstract_inverted_index.responses | 193 |
| abstract_inverted_index.therapies | 13 |
| abstract_inverted_index.wild-type | 143 |
| abstract_inverted_index.(SARS)-CoV | 56 |
| abstract_inverted_index.ADP-ribose | 31 |
| abstract_inverted_index.SARS-CoV-1 | 258 |
| abstract_inverted_index.SARS-CoV-2 | 116, 137, 171, 221, 231, 277, 292, 309 |
| abstract_inverted_index.additional | 357 |
| abstract_inverted_index.antivirals | 286 |
| abstract_inverted_index.artificial | 89 |
| abstract_inverted_index.containing | 92, 310 |
| abstract_inverted_index.contribute | 196 |
| abstract_inverted_index.difference | 127 |
| abstract_inverted_index.expression | 182 |
| abstract_inverted_index.functions, | 354 |
| abstract_inverted_index.imperative | 66 |
| abstract_inverted_index.indicating | 124, 188 |
| abstract_inverted_index.infection, | 159 |
| abstract_inverted_index.infection. | 151 |
| abstract_inverted_index.infectious | 108 |
| abstract_inverted_index.interferon | 176 |
| abstract_inverted_index.morbidity. | 169 |
| abstract_inverted_index.production | 266 |
| abstract_inverted_index.replicated | 139 |
| abstract_inverted_index.replicates | 295, 315 |
| abstract_inverted_index.therapies. | 361 |
| abstract_inverted_index.understand | 68, 290 |
| abstract_inverted_index.virulence. | 57 |
| abstract_inverted_index.SARS-CoV-2, | 244 |
| abstract_inverted_index.SARS-CoV-2. | 101 |
| abstract_inverted_index.chromosomes | 90 |
| abstract_inverted_index.coronavirus | 2, 360 |
| abstract_inverted_index.demonstrate | 215 |
| abstract_inverted_index.development | 275, 281 |
| abstract_inverted_index.inhibitors, | 350 |
| abstract_inverted_index.inhibitors. | 279 |
| abstract_inverted_index.macrodomain | 249 |
| abstract_inverted_index.recombinant | 37, 87 |
| abstract_inverted_index.replication | 76 |
| abstract_inverted_index.requirement | 130 |
| abstract_inverted_index.respiratory | 54 |
| abstract_inverted_index.significant | 337 |
| abstract_inverted_index.susceptible | 149 |
| abstract_inverted_index.Furthermore, | 136 |
| abstract_inverted_index.SIGNIFICANCE | 240 |
| abstract_inverted_index.attenuation. | 199 |
| abstract_inverted_index.inflammatory | 209 |
| abstract_inverted_index.macrodomain, | 25 |
| abstract_inverted_index.neutrophils. | 212 |
| abstract_inverted_index.pathogenesis | 78, 232 |
| abstract_inverted_index.replication, | 222 |
| abstract_inverted_index.pathogenesis, | 270 |
| abstract_inverted_index.transfection, | 123 |
| abstract_inverted_index.ADP-ribosylation. | 254 |
| abstract_inverted_index.ADP-ribosylhydrolase | 29 |
| abstract_inverted_index.interferon-stimulated | 179 |
| cited_by_percentile_year.max | 99 |
| cited_by_percentile_year.min | 91 |
| corresponding_author_ids | https://openalex.org/A5078895477, https://openalex.org/A5023943993 |
| countries_distinct_count | 2 |
| institutions_distinct_count | 14 |
| corresponding_institution_ids | https://openalex.org/I115475287, https://openalex.org/I146416000 |
| sustainable_development_goals[0].id | https://metadata.un.org/sdg/3 |
| sustainable_development_goals[0].score | 0.8100000023841858 |
| sustainable_development_goals[0].display_name | Good health and well-being |
| citation_normalized_percentile.value | 0.89101847 |
| citation_normalized_percentile.is_in_top_1_percent | False |
| citation_normalized_percentile.is_in_top_10_percent | True |