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Structure of a functional cap-binding domain in Rift Valley fever virus L protein
Rift Valley fever virus (RVFV) belongs to the family of Phenuiviridae within the order of Bunyavirales. The virus may cause fatal disease both in livestock and humans, and therefore, is of great economical and public health relevance. In analogy to the influenza virus polymerase complex, the bunyavi...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Public Library of Science
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6555543/ https://www.ncbi.nlm.nih.gov/pubmed/31136637 http://dx.doi.org/10.1371/journal.ppat.1007829 |
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author | Gogrefe, Nadja Reindl, Sophia Günther, Stephan Rosenthal, Maria |
author_facet | Gogrefe, Nadja Reindl, Sophia Günther, Stephan Rosenthal, Maria |
author_sort | Gogrefe, Nadja |
collection | PubMed |
description | Rift Valley fever virus (RVFV) belongs to the family of Phenuiviridae within the order of Bunyavirales. The virus may cause fatal disease both in livestock and humans, and therefore, is of great economical and public health relevance. In analogy to the influenza virus polymerase complex, the bunyavirus L protein is assumed to bind to and cleave off cap structures of cellular mRNAs to prime viral transcription. However, even though the presence of an endonuclease in the N-terminal domain of the L protein has been demonstrated for several bunyaviruses, there is no evidence for a cap-binding site within the L protein. We solved the structure of a C-terminal 117 amino acid-long domain of the RVFV L protein by X-ray crystallography. The overall fold of the domain shows high similarity to influenza virus PB2 cap-binding domain and the putative non-functional cap-binding domain of reptarenaviruses. Upon co-crystallization with m(7)GTP, we detected the cap-analogue bound between two aromatic side chains as it has been described for other cap-binding proteins. We observed weak but specific interaction with m(7)GTP rather than GTP in vitro using isothermal titration calorimetry. The importance of m(7)GTP-binding residues for viral transcription was validated using a RVFV minigenome system. In summary, we provide structural and functional evidence for a cap-binding site located within the L protein of a virus from the Bunyavirales order. |
format | Online Article Text |
id | pubmed-6555543 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-65555432019-06-17 Structure of a functional cap-binding domain in Rift Valley fever virus L protein Gogrefe, Nadja Reindl, Sophia Günther, Stephan Rosenthal, Maria PLoS Pathog Research Article Rift Valley fever virus (RVFV) belongs to the family of Phenuiviridae within the order of Bunyavirales. The virus may cause fatal disease both in livestock and humans, and therefore, is of great economical and public health relevance. In analogy to the influenza virus polymerase complex, the bunyavirus L protein is assumed to bind to and cleave off cap structures of cellular mRNAs to prime viral transcription. However, even though the presence of an endonuclease in the N-terminal domain of the L protein has been demonstrated for several bunyaviruses, there is no evidence for a cap-binding site within the L protein. We solved the structure of a C-terminal 117 amino acid-long domain of the RVFV L protein by X-ray crystallography. The overall fold of the domain shows high similarity to influenza virus PB2 cap-binding domain and the putative non-functional cap-binding domain of reptarenaviruses. Upon co-crystallization with m(7)GTP, we detected the cap-analogue bound between two aromatic side chains as it has been described for other cap-binding proteins. We observed weak but specific interaction with m(7)GTP rather than GTP in vitro using isothermal titration calorimetry. The importance of m(7)GTP-binding residues for viral transcription was validated using a RVFV minigenome system. In summary, we provide structural and functional evidence for a cap-binding site located within the L protein of a virus from the Bunyavirales order. Public Library of Science 2019-05-28 /pmc/articles/PMC6555543/ /pubmed/31136637 http://dx.doi.org/10.1371/journal.ppat.1007829 Text en © 2019 Gogrefe 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 (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article Gogrefe, Nadja Reindl, Sophia Günther, Stephan Rosenthal, Maria Structure of a functional cap-binding domain in Rift Valley fever virus L protein |
title | Structure of a functional cap-binding domain in Rift Valley fever virus L protein |
title_full | Structure of a functional cap-binding domain in Rift Valley fever virus L protein |
title_fullStr | Structure of a functional cap-binding domain in Rift Valley fever virus L protein |
title_full_unstemmed | Structure of a functional cap-binding domain in Rift Valley fever virus L protein |
title_short | Structure of a functional cap-binding domain in Rift Valley fever virus L protein |
title_sort | structure of a functional cap-binding domain in rift valley fever virus l protein |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6555543/ https://www.ncbi.nlm.nih.gov/pubmed/31136637 http://dx.doi.org/10.1371/journal.ppat.1007829 |
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