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Structural Basis for the dsRNA Specificity of the Lassa Virus NP Exonuclease

Lassa virus causes hemorrhagic fever characterized by immunosuppression. The nucleoprotein of Lassa virus, termed NP, binds the viral genome. It also has an additional enzymatic activity as an exonuclease that specifically digests double-stranded RNA (dsRNA). dsRNA is a strong signal to the innate i...

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Autores principales: Hastie, Kathryn M., King, Liam B., Zandonatti, Michelle A., Saphire, Erica Ollmann
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3429428/
https://www.ncbi.nlm.nih.gov/pubmed/22937163
http://dx.doi.org/10.1371/journal.pone.0044211
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author Hastie, Kathryn M.
King, Liam B.
Zandonatti, Michelle A.
Saphire, Erica Ollmann
author_facet Hastie, Kathryn M.
King, Liam B.
Zandonatti, Michelle A.
Saphire, Erica Ollmann
author_sort Hastie, Kathryn M.
collection PubMed
description Lassa virus causes hemorrhagic fever characterized by immunosuppression. The nucleoprotein of Lassa virus, termed NP, binds the viral genome. It also has an additional enzymatic activity as an exonuclease that specifically digests double-stranded RNA (dsRNA). dsRNA is a strong signal to the innate immune system of viral infection. Digestion of dsRNA by the NP exonuclease activity appears to cause suppression of innate immune signaling in the infected cell. Although the fold of the NP enzyme is conserved and the active site completely conserved with other exonucleases in its DEDDh family, NP is atypical among exonucleases in its preference for dsRNA and its strict specificity for one substrate. Here, we present the crystal structure of Lassa virus NP in complex with dsRNA. We find that unlike the exonuclease in Klenow fragment, the double-stranded nucleic acid in complex with Lassa NP remains base-paired instead of splitting, and that binding of the paired complementary strand is achieved by “relocation” of a basic loop motif from its typical exonuclease position. Further, we find that just one single glycine that contacts the substrate strand and one single tyrosine that stacks with a base of the complementary, non-substrate strand are responsible for the unique substrate specificity. This work thus provides templates for development of antiviral drugs that would be specific for viral, rather than host exonucleases of similar fold and active site, and illustrates how a very few amino acid changes confer alternate specificity and biological phenotype to an enzyme.
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spelling pubmed-34294282012-08-30 Structural Basis for the dsRNA Specificity of the Lassa Virus NP Exonuclease Hastie, Kathryn M. King, Liam B. Zandonatti, Michelle A. Saphire, Erica Ollmann PLoS One Research Article Lassa virus causes hemorrhagic fever characterized by immunosuppression. The nucleoprotein of Lassa virus, termed NP, binds the viral genome. It also has an additional enzymatic activity as an exonuclease that specifically digests double-stranded RNA (dsRNA). dsRNA is a strong signal to the innate immune system of viral infection. Digestion of dsRNA by the NP exonuclease activity appears to cause suppression of innate immune signaling in the infected cell. Although the fold of the NP enzyme is conserved and the active site completely conserved with other exonucleases in its DEDDh family, NP is atypical among exonucleases in its preference for dsRNA and its strict specificity for one substrate. Here, we present the crystal structure of Lassa virus NP in complex with dsRNA. We find that unlike the exonuclease in Klenow fragment, the double-stranded nucleic acid in complex with Lassa NP remains base-paired instead of splitting, and that binding of the paired complementary strand is achieved by “relocation” of a basic loop motif from its typical exonuclease position. Further, we find that just one single glycine that contacts the substrate strand and one single tyrosine that stacks with a base of the complementary, non-substrate strand are responsible for the unique substrate specificity. This work thus provides templates for development of antiviral drugs that would be specific for viral, rather than host exonucleases of similar fold and active site, and illustrates how a very few amino acid changes confer alternate specificity and biological phenotype to an enzyme. Public Library of Science 2012-08-28 /pmc/articles/PMC3429428/ /pubmed/22937163 http://dx.doi.org/10.1371/journal.pone.0044211 Text en © 2012 Hastie, 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
Hastie, Kathryn M.
King, Liam B.
Zandonatti, Michelle A.
Saphire, Erica Ollmann
Structural Basis for the dsRNA Specificity of the Lassa Virus NP Exonuclease
title Structural Basis for the dsRNA Specificity of the Lassa Virus NP Exonuclease
title_full Structural Basis for the dsRNA Specificity of the Lassa Virus NP Exonuclease
title_fullStr Structural Basis for the dsRNA Specificity of the Lassa Virus NP Exonuclease
title_full_unstemmed Structural Basis for the dsRNA Specificity of the Lassa Virus NP Exonuclease
title_short Structural Basis for the dsRNA Specificity of the Lassa Virus NP Exonuclease
title_sort structural basis for the dsrna specificity of the lassa virus np exonuclease
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3429428/
https://www.ncbi.nlm.nih.gov/pubmed/22937163
http://dx.doi.org/10.1371/journal.pone.0044211
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