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The N-Terminus of the RNA Polymerase from Infectious Pancreatic Necrosis Virus Is the Determinant of Genome Attachment
The RNA-dependent RNA polymerase VP1 of infectious pancreatic necrosis virus (IPNV) is a single polypeptide responsible for both viral RNA transcription and genome replication. Sequence analysis identifies IPNV VP1 as having an unusual active site topology. We have purified, crystallized and solved...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2011
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3121795/ https://www.ncbi.nlm.nih.gov/pubmed/21731487 http://dx.doi.org/10.1371/journal.ppat.1002085 |
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author | Graham, Stephen C. Sarin, L. Peter Bahar, Mohammad W. Myers, Reg A. Stuart, David I. Bamford, Dennis H. Grimes, Jonathan M. |
author_facet | Graham, Stephen C. Sarin, L. Peter Bahar, Mohammad W. Myers, Reg A. Stuart, David I. Bamford, Dennis H. Grimes, Jonathan M. |
author_sort | Graham, Stephen C. |
collection | PubMed |
description | The RNA-dependent RNA polymerase VP1 of infectious pancreatic necrosis virus (IPNV) is a single polypeptide responsible for both viral RNA transcription and genome replication. Sequence analysis identifies IPNV VP1 as having an unusual active site topology. We have purified, crystallized and solved the structure of IPNV VP1 to 2.3 Å resolution in its apo form and at 2.2 Å resolution bound to the catalytically-activating metal magnesium. We find that recombinantly-expressed VP1 is highly active for RNA transcription and replication, yielding both free and polymerase-attached RNA products. IPNV VP1 also possesses terminal (deoxy)nucleotide transferase, RNA-dependent DNA polymerase (reverse transcriptase) and template-independent self-guanylylation activity. The N-terminus of VP1 interacts with the active-site cleft and we show that the N-terminal serine residue is required for formation of covalent RNA∶polymerase complexes, providing a mechanism for the genesis of viral genome∶polymerase complexes observed in vivo. |
format | Online Article Text |
id | pubmed-3121795 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2011 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-31217952011-06-30 The N-Terminus of the RNA Polymerase from Infectious Pancreatic Necrosis Virus Is the Determinant of Genome Attachment Graham, Stephen C. Sarin, L. Peter Bahar, Mohammad W. Myers, Reg A. Stuart, David I. Bamford, Dennis H. Grimes, Jonathan M. PLoS Pathog Research Article The RNA-dependent RNA polymerase VP1 of infectious pancreatic necrosis virus (IPNV) is a single polypeptide responsible for both viral RNA transcription and genome replication. Sequence analysis identifies IPNV VP1 as having an unusual active site topology. We have purified, crystallized and solved the structure of IPNV VP1 to 2.3 Å resolution in its apo form and at 2.2 Å resolution bound to the catalytically-activating metal magnesium. We find that recombinantly-expressed VP1 is highly active for RNA transcription and replication, yielding both free and polymerase-attached RNA products. IPNV VP1 also possesses terminal (deoxy)nucleotide transferase, RNA-dependent DNA polymerase (reverse transcriptase) and template-independent self-guanylylation activity. The N-terminus of VP1 interacts with the active-site cleft and we show that the N-terminal serine residue is required for formation of covalent RNA∶polymerase complexes, providing a mechanism for the genesis of viral genome∶polymerase complexes observed in vivo. Public Library of Science 2011-06-23 /pmc/articles/PMC3121795/ /pubmed/21731487 http://dx.doi.org/10.1371/journal.ppat.1002085 Text en Graham et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Graham, Stephen C. Sarin, L. Peter Bahar, Mohammad W. Myers, Reg A. Stuart, David I. Bamford, Dennis H. Grimes, Jonathan M. The N-Terminus of the RNA Polymerase from Infectious Pancreatic Necrosis Virus Is the Determinant of Genome Attachment |
title | The N-Terminus of the RNA Polymerase from Infectious Pancreatic Necrosis Virus Is the Determinant of Genome Attachment |
title_full | The N-Terminus of the RNA Polymerase from Infectious Pancreatic Necrosis Virus Is the Determinant of Genome Attachment |
title_fullStr | The N-Terminus of the RNA Polymerase from Infectious Pancreatic Necrosis Virus Is the Determinant of Genome Attachment |
title_full_unstemmed | The N-Terminus of the RNA Polymerase from Infectious Pancreatic Necrosis Virus Is the Determinant of Genome Attachment |
title_short | The N-Terminus of the RNA Polymerase from Infectious Pancreatic Necrosis Virus Is the Determinant of Genome Attachment |
title_sort | n-terminus of the rna polymerase from infectious pancreatic necrosis virus is the determinant of genome attachment |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3121795/ https://www.ncbi.nlm.nih.gov/pubmed/21731487 http://dx.doi.org/10.1371/journal.ppat.1002085 |
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