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Conserved Endonuclease Function of Hantavirus L Polymerase

Hantaviruses are important emerging pathogens belonging to the Bunyaviridae family. Like other segmented negative strand RNA viruses, the RNA-dependent RNA polymerase (RdRp) also known as L protein of hantaviruses lacks an intrinsic “capping activity”. Hantaviruses therefore employ a “cap snatching”...

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Autores principales: Rothenberger, Sylvia, Torriani, Giulia, Johansson, Maria U., Kunz, Stefan, Engler, Olivier
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4885073/
https://www.ncbi.nlm.nih.gov/pubmed/27144576
http://dx.doi.org/10.3390/v8050108
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author Rothenberger, Sylvia
Torriani, Giulia
Johansson, Maria U.
Kunz, Stefan
Engler, Olivier
author_facet Rothenberger, Sylvia
Torriani, Giulia
Johansson, Maria U.
Kunz, Stefan
Engler, Olivier
author_sort Rothenberger, Sylvia
collection PubMed
description Hantaviruses are important emerging pathogens belonging to the Bunyaviridae family. Like other segmented negative strand RNA viruses, the RNA-dependent RNA polymerase (RdRp) also known as L protein of hantaviruses lacks an intrinsic “capping activity”. Hantaviruses therefore employ a “cap snatching” strategy acquiring short 5′ RNA sequences bearing 5′cap structures by endonucleolytic cleavage from host cell transcripts. The viral endonuclease activity implicated in cap snatching of hantaviruses has been mapped to the N-terminal domain of the L protein. Using a combination of molecular modeling and structure–function analysis we confirm and extend these findings providing evidence for high conservation of the L endonuclease between Old and New World hantaviruses. Recombinant hantavirus L endonuclease showed catalytic activity and a defined cation preference shared by other viral endonucleases. Based on the previously reported remarkably high activity of hantavirus L endonuclease, we established a cell-based assay for the hantavirus endonuclase function. The robustness of the assay and its high-throughput compatible format makes it suitable for small molecule drug screens to identify novel inhibitors of hantavirus endonuclease. Based on the high degree of similarity to RdRp endonucleases, some candidate inhibitors may be broadly active against hantaviruses and other emerging human pathogenic Bunyaviruses.
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spelling pubmed-48850732016-05-31 Conserved Endonuclease Function of Hantavirus L Polymerase Rothenberger, Sylvia Torriani, Giulia Johansson, Maria U. Kunz, Stefan Engler, Olivier Viruses Article Hantaviruses are important emerging pathogens belonging to the Bunyaviridae family. Like other segmented negative strand RNA viruses, the RNA-dependent RNA polymerase (RdRp) also known as L protein of hantaviruses lacks an intrinsic “capping activity”. Hantaviruses therefore employ a “cap snatching” strategy acquiring short 5′ RNA sequences bearing 5′cap structures by endonucleolytic cleavage from host cell transcripts. The viral endonuclease activity implicated in cap snatching of hantaviruses has been mapped to the N-terminal domain of the L protein. Using a combination of molecular modeling and structure–function analysis we confirm and extend these findings providing evidence for high conservation of the L endonuclease between Old and New World hantaviruses. Recombinant hantavirus L endonuclease showed catalytic activity and a defined cation preference shared by other viral endonucleases. Based on the previously reported remarkably high activity of hantavirus L endonuclease, we established a cell-based assay for the hantavirus endonuclase function. The robustness of the assay and its high-throughput compatible format makes it suitable for small molecule drug screens to identify novel inhibitors of hantavirus endonuclease. Based on the high degree of similarity to RdRp endonucleases, some candidate inhibitors may be broadly active against hantaviruses and other emerging human pathogenic Bunyaviruses. MDPI 2016-05-02 /pmc/articles/PMC4885073/ /pubmed/27144576 http://dx.doi.org/10.3390/v8050108 Text en © 2016 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC-BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Rothenberger, Sylvia
Torriani, Giulia
Johansson, Maria U.
Kunz, Stefan
Engler, Olivier
Conserved Endonuclease Function of Hantavirus L Polymerase
title Conserved Endonuclease Function of Hantavirus L Polymerase
title_full Conserved Endonuclease Function of Hantavirus L Polymerase
title_fullStr Conserved Endonuclease Function of Hantavirus L Polymerase
title_full_unstemmed Conserved Endonuclease Function of Hantavirus L Polymerase
title_short Conserved Endonuclease Function of Hantavirus L Polymerase
title_sort conserved endonuclease function of hantavirus l polymerase
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4885073/
https://www.ncbi.nlm.nih.gov/pubmed/27144576
http://dx.doi.org/10.3390/v8050108
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