Structure of the recombinant RNA polymerase from African Swine Fever Virus
Abstract African Swine Fever Virus is a Nucleo-Cytoplasmic Large DNA Virus that causes an incurable haemorrhagic fever in pigs with a high impact on global food security. ASFV replicates in the cytoplasm of the infected cell and encodes its own transcription machinery that is independent of cellular...
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2024-02-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-024-45842-7 |
| _version_ | 1797274010390626304 |
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| author | Simona Pilotto Michal Sýkora Gwenny Cackett Christopher Dulson Finn Werner |
| author_facet | Simona Pilotto Michal Sýkora Gwenny Cackett Christopher Dulson Finn Werner |
| author_sort | Simona Pilotto |
| collection | DOAJ |
| description | Abstract African Swine Fever Virus is a Nucleo-Cytoplasmic Large DNA Virus that causes an incurable haemorrhagic fever in pigs with a high impact on global food security. ASFV replicates in the cytoplasm of the infected cell and encodes its own transcription machinery that is independent of cellular factors, however, not much is known about how this system works at a molecular level. Here, we present methods to produce recombinant ASFV RNA polymerase, functional assays to screen for inhibitors, and high-resolution cryo-electron microscopy structures of the ASFV RNAP in different conformational states. The ASFV RNAP bears a striking resemblance to RNAPII with bona fide homologues of nine of its twelve subunits. Key differences include the fusion of the ASFV assembly platform subunits RPB3 and RPB11, and an unusual C-terminal domain of the stalk subunit vRPB7 that is related to the eukaryotic mRNA cap 2´-O-methyltransferase 1. Despite the high degree of structural conservation with cellular RNA polymerases, the ASFV RNAP is resistant to the inhibitors rifampicin and alpha-amanitin. The cryo-EM structures and fully recombinant RNAP system together provide an important tool for the design, development, and screening of antiviral drugs in a low biosafety containment environment. |
| first_indexed | 2024-03-07T14:52:16Z |
| format | Article |
| id | doaj.art-21d5307af46c4ea2820b8a8886e4a5d0 |
| institution | Directory Open Access Journal |
| issn | 2041-1723 |
| language | English |
| last_indexed | 2024-03-07T14:52:16Z |
| publishDate | 2024-02-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj.art-21d5307af46c4ea2820b8a8886e4a5d02024-03-05T19:37:33ZengNature PortfolioNature Communications2041-17232024-02-0115111510.1038/s41467-024-45842-7Structure of the recombinant RNA polymerase from African Swine Fever VirusSimona Pilotto0Michal Sýkora1Gwenny Cackett2Christopher Dulson3Finn Werner4Institute for Structural and Molecular Biology, Division of Biosciences, University College London, Gower StreetInstitute for Structural and Molecular Biology, Division of Biosciences, University College London, Gower StreetInstitute for Structural and Molecular Biology, Division of Biosciences, University College London, Gower StreetInstitute for Structural and Molecular Biology, Division of Biosciences, University College London, Gower StreetInstitute for Structural and Molecular Biology, Division of Biosciences, University College London, Gower StreetAbstract African Swine Fever Virus is a Nucleo-Cytoplasmic Large DNA Virus that causes an incurable haemorrhagic fever in pigs with a high impact on global food security. ASFV replicates in the cytoplasm of the infected cell and encodes its own transcription machinery that is independent of cellular factors, however, not much is known about how this system works at a molecular level. Here, we present methods to produce recombinant ASFV RNA polymerase, functional assays to screen for inhibitors, and high-resolution cryo-electron microscopy structures of the ASFV RNAP in different conformational states. The ASFV RNAP bears a striking resemblance to RNAPII with bona fide homologues of nine of its twelve subunits. Key differences include the fusion of the ASFV assembly platform subunits RPB3 and RPB11, and an unusual C-terminal domain of the stalk subunit vRPB7 that is related to the eukaryotic mRNA cap 2´-O-methyltransferase 1. Despite the high degree of structural conservation with cellular RNA polymerases, the ASFV RNAP is resistant to the inhibitors rifampicin and alpha-amanitin. The cryo-EM structures and fully recombinant RNAP system together provide an important tool for the design, development, and screening of antiviral drugs in a low biosafety containment environment.https://doi.org/10.1038/s41467-024-45842-7 |
| spellingShingle | Simona Pilotto Michal Sýkora Gwenny Cackett Christopher Dulson Finn Werner Structure of the recombinant RNA polymerase from African Swine Fever Virus Nature Communications |
| title | Structure of the recombinant RNA polymerase from African Swine Fever Virus |
| title_full | Structure of the recombinant RNA polymerase from African Swine Fever Virus |
| title_fullStr | Structure of the recombinant RNA polymerase from African Swine Fever Virus |
| title_full_unstemmed | Structure of the recombinant RNA polymerase from African Swine Fever Virus |
| title_short | Structure of the recombinant RNA polymerase from African Swine Fever Virus |
| title_sort | structure of the recombinant rna polymerase from african swine fever virus |
| url | https://doi.org/10.1038/s41467-024-45842-7 |
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