Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Graham, Stephen C., Sarin, L. Peter, Bahar, Mohammad W., Myers, Reg A., Stuart, David I., Bamford, Dennis H., Grimes, Jonathan M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2011
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
_version_ 1782206872905842688
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
work_keys_str_mv AT grahamstephenc thenterminusofthernapolymerasefrominfectiouspancreaticnecrosisvirusisthedeterminantofgenomeattachment
AT sarinlpeter thenterminusofthernapolymerasefrominfectiouspancreaticnecrosisvirusisthedeterminantofgenomeattachment
AT baharmohammadw thenterminusofthernapolymerasefrominfectiouspancreaticnecrosisvirusisthedeterminantofgenomeattachment
AT myersrega thenterminusofthernapolymerasefrominfectiouspancreaticnecrosisvirusisthedeterminantofgenomeattachment
AT stuartdavidi thenterminusofthernapolymerasefrominfectiouspancreaticnecrosisvirusisthedeterminantofgenomeattachment
AT bamforddennish thenterminusofthernapolymerasefrominfectiouspancreaticnecrosisvirusisthedeterminantofgenomeattachment
AT grimesjonathanm thenterminusofthernapolymerasefrominfectiouspancreaticnecrosisvirusisthedeterminantofgenomeattachment
AT grahamstephenc nterminusofthernapolymerasefrominfectiouspancreaticnecrosisvirusisthedeterminantofgenomeattachment
AT sarinlpeter nterminusofthernapolymerasefrominfectiouspancreaticnecrosisvirusisthedeterminantofgenomeattachment
AT baharmohammadw nterminusofthernapolymerasefrominfectiouspancreaticnecrosisvirusisthedeterminantofgenomeattachment
AT myersrega nterminusofthernapolymerasefrominfectiouspancreaticnecrosisvirusisthedeterminantofgenomeattachment
AT stuartdavidi nterminusofthernapolymerasefrominfectiouspancreaticnecrosisvirusisthedeterminantofgenomeattachment
AT bamforddennish nterminusofthernapolymerasefrominfectiouspancreaticnecrosisvirusisthedeterminantofgenomeattachment
AT grimesjonathanm nterminusofthernapolymerasefrominfectiouspancreaticnecrosisvirusisthedeterminantofgenomeattachment