The Dengue Virus NS5 Protein Intrudes in the Cellular Spliceosome and Modulates Splicing

Dengue virus NS5 protein plays multiple functions in the cytoplasm of infected cells, enabling viral RNA replication and counteracting host antiviral responses. Here, we demonstrate a novel function of NS5 in the nucleus where it interferes with cellular splicing. Using global proteomic analysis of...

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Autores principales:	De Maio, F.A., Risso, G., Iglesias, N.G., Shah, P., Pozzi, B., Gebhard, L.G., Mammi, P., Mancini, E., Yanovsky, M.J., Andino, R., Krogan, N., Srebrow, A., Gamarnik, A.V.
Formato:	JOUR
Materias:	complementary DNA cystic fibrosis transmembrane conductance regulator monoclonal antibody nonstructural protein 5 polyclonal antibody protein bcl x small nuclear ribonucleoprotein STAT2 protein NS5 protein, dengue virus viral protein A549 cell line alternative RNA splicing Article cell fractionation controlled study dengue Dengue virus gene expression regulation genetic transfection hepatocellular carcinoma cell line high throughput sequencing host pathogen interaction human human cell immunofluorescence test protein localization protein protein interaction protein purification proteomics real time polymerase chain reaction RNA processing RNAi therapeutics spliceosome virus recombinant virus replication Western blotting animal cell line fluorescent antibody technique genetics host parasite interaction metabolism pathogenicity physiology polymerase chain reaction RNA splicing virology Animals Cell Line Dengue Dengue Virus Fluorescent Antibody Technique High-Throughput Nucleotide Sequencing Host-Parasite Interactions Humans Polymerase Chain Reaction Ribonucleoprotein, U5 Small Nuclear RNA Splicing Spliceosomes Transfection Viral Nonstructural Proteins
Acceso en línea:	http://hdl.handle.net/20.500.12110/paper_15537366_v12_n8_p_DeMaio
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

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spelling	todo:paper_15537366_v12_n8_p_DeMaio2023-10-03T16:25:31Z The Dengue Virus NS5 Protein Intrudes in the Cellular Spliceosome and Modulates Splicing De Maio, F.A. Risso, G. Iglesias, N.G. Shah, P. Pozzi, B. Gebhard, L.G. Mammi, P. Mancini, E. Yanovsky, M.J. Andino, R. Krogan, N. Srebrow, A. Gamarnik, A.V. complementary DNA cystic fibrosis transmembrane conductance regulator monoclonal antibody nonstructural protein 5 polyclonal antibody protein bcl x small nuclear ribonucleoprotein STAT2 protein NS5 protein, dengue virus small nuclear ribonucleoprotein viral protein A549 cell line alternative RNA splicing Article cell fractionation controlled study dengue Dengue virus gene expression regulation genetic transfection hepatocellular carcinoma cell line high throughput sequencing host pathogen interaction human human cell immunofluorescence test protein localization protein protein interaction protein purification proteomics real time polymerase chain reaction RNA processing RNAi therapeutics spliceosome virus recombinant virus replication Western blotting animal cell line fluorescent antibody technique genetics host parasite interaction metabolism pathogenicity physiology polymerase chain reaction RNA splicing spliceosome virology Animals Cell Line Dengue Dengue Virus Fluorescent Antibody Technique High-Throughput Nucleotide Sequencing Host-Parasite Interactions Humans Polymerase Chain Reaction Ribonucleoprotein, U5 Small Nuclear RNA Splicing Spliceosomes Transfection Viral Nonstructural Proteins Dengue virus NS5 protein plays multiple functions in the cytoplasm of infected cells, enabling viral RNA replication and counteracting host antiviral responses. Here, we demonstrate a novel function of NS5 in the nucleus where it interferes with cellular splicing. Using global proteomic analysis of infected cells together with functional studies, we found that NS5 binds spliceosome complexes and modulates endogenous splicing as well as minigene-derived alternative splicing patterns. In particular, we show that NS5 alone, or in the context of viral infection, interacts with core components of the U5 snRNP particle, CD2BP2 and DDX23, alters the inclusion/exclusion ratio of alternative splicing events, and changes mRNA isoform abundance of known antiviral factors. Interestingly, a genome wide transcriptome analysis, using recently developed bioinformatics tools, revealed an increase of intron retention upon dengue virus infection, and viral replication was improved by silencing specific U5 components. Different mechanistic studies indicate that binding of NS5 to the spliceosome reduces the efficiency of pre-mRNA processing, independently of NS5 enzymatic activities. We propose that NS5 binding to U5 snRNP proteins hijacks the splicing machinery resulting in a less restrictive environment for viral replication. © 2016 De Maio et al. Fil:Risso, G. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Pozzi, B. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Yanovsky, M.J. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Srebrow, A. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_15537366_v12_n8_p_DeMaio
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic	complementary DNA cystic fibrosis transmembrane conductance regulator monoclonal antibody nonstructural protein 5 polyclonal antibody protein bcl x small nuclear ribonucleoprotein STAT2 protein NS5 protein, dengue virus small nuclear ribonucleoprotein viral protein A549 cell line alternative RNA splicing Article cell fractionation controlled study dengue Dengue virus gene expression regulation genetic transfection hepatocellular carcinoma cell line high throughput sequencing host pathogen interaction human human cell immunofluorescence test protein localization protein protein interaction protein purification proteomics real time polymerase chain reaction RNA processing RNAi therapeutics spliceosome virus recombinant virus replication Western blotting animal cell line fluorescent antibody technique genetics host parasite interaction metabolism pathogenicity physiology polymerase chain reaction RNA splicing spliceosome virology Animals Cell Line Dengue Dengue Virus Fluorescent Antibody Technique High-Throughput Nucleotide Sequencing Host-Parasite Interactions Humans Polymerase Chain Reaction Ribonucleoprotein, U5 Small Nuclear RNA Splicing Spliceosomes Transfection Viral Nonstructural Proteins
spellingShingle	complementary DNA cystic fibrosis transmembrane conductance regulator monoclonal antibody nonstructural protein 5 polyclonal antibody protein bcl x small nuclear ribonucleoprotein STAT2 protein NS5 protein, dengue virus small nuclear ribonucleoprotein viral protein A549 cell line alternative RNA splicing Article cell fractionation controlled study dengue Dengue virus gene expression regulation genetic transfection hepatocellular carcinoma cell line high throughput sequencing host pathogen interaction human human cell immunofluorescence test protein localization protein protein interaction protein purification proteomics real time polymerase chain reaction RNA processing RNAi therapeutics spliceosome virus recombinant virus replication Western blotting animal cell line fluorescent antibody technique genetics host parasite interaction metabolism pathogenicity physiology polymerase chain reaction RNA splicing spliceosome virology Animals Cell Line Dengue Dengue Virus Fluorescent Antibody Technique High-Throughput Nucleotide Sequencing Host-Parasite Interactions Humans Polymerase Chain Reaction Ribonucleoprotein, U5 Small Nuclear RNA Splicing Spliceosomes Transfection Viral Nonstructural Proteins De Maio, F.A. Risso, G. Iglesias, N.G. Shah, P. Pozzi, B. Gebhard, L.G. Mammi, P. Mancini, E. Yanovsky, M.J. Andino, R. Krogan, N. Srebrow, A. Gamarnik, A.V. The Dengue Virus NS5 Protein Intrudes in the Cellular Spliceosome and Modulates Splicing
topic_facet	complementary DNA cystic fibrosis transmembrane conductance regulator monoclonal antibody nonstructural protein 5 polyclonal antibody protein bcl x small nuclear ribonucleoprotein STAT2 protein NS5 protein, dengue virus small nuclear ribonucleoprotein viral protein A549 cell line alternative RNA splicing Article cell fractionation controlled study dengue Dengue virus gene expression regulation genetic transfection hepatocellular carcinoma cell line high throughput sequencing host pathogen interaction human human cell immunofluorescence test protein localization protein protein interaction protein purification proteomics real time polymerase chain reaction RNA processing RNAi therapeutics spliceosome virus recombinant virus replication Western blotting animal cell line fluorescent antibody technique genetics host parasite interaction metabolism pathogenicity physiology polymerase chain reaction RNA splicing spliceosome virology Animals Cell Line Dengue Dengue Virus Fluorescent Antibody Technique High-Throughput Nucleotide Sequencing Host-Parasite Interactions Humans Polymerase Chain Reaction Ribonucleoprotein, U5 Small Nuclear RNA Splicing Spliceosomes Transfection Viral Nonstructural Proteins
description	Dengue virus NS5 protein plays multiple functions in the cytoplasm of infected cells, enabling viral RNA replication and counteracting host antiviral responses. Here, we demonstrate a novel function of NS5 in the nucleus where it interferes with cellular splicing. Using global proteomic analysis of infected cells together with functional studies, we found that NS5 binds spliceosome complexes and modulates endogenous splicing as well as minigene-derived alternative splicing patterns. In particular, we show that NS5 alone, or in the context of viral infection, interacts with core components of the U5 snRNP particle, CD2BP2 and DDX23, alters the inclusion/exclusion ratio of alternative splicing events, and changes mRNA isoform abundance of known antiviral factors. Interestingly, a genome wide transcriptome analysis, using recently developed bioinformatics tools, revealed an increase of intron retention upon dengue virus infection, and viral replication was improved by silencing specific U5 components. Different mechanistic studies indicate that binding of NS5 to the spliceosome reduces the efficiency of pre-mRNA processing, independently of NS5 enzymatic activities. We propose that NS5 binding to U5 snRNP proteins hijacks the splicing machinery resulting in a less restrictive environment for viral replication. © 2016 De Maio et al.
format	JOUR
author	De Maio, F.A. Risso, G. Iglesias, N.G. Shah, P. Pozzi, B. Gebhard, L.G. Mammi, P. Mancini, E. Yanovsky, M.J. Andino, R. Krogan, N. Srebrow, A. Gamarnik, A.V.
author_facet	De Maio, F.A. Risso, G. Iglesias, N.G. Shah, P. Pozzi, B. Gebhard, L.G. Mammi, P. Mancini, E. Yanovsky, M.J. Andino, R. Krogan, N. Srebrow, A. Gamarnik, A.V.
author_sort	De Maio, F.A.
title	The Dengue Virus NS5 Protein Intrudes in the Cellular Spliceosome and Modulates Splicing
title_short	The Dengue Virus NS5 Protein Intrudes in the Cellular Spliceosome and Modulates Splicing
title_full	The Dengue Virus NS5 Protein Intrudes in the Cellular Spliceosome and Modulates Splicing
title_fullStr	The Dengue Virus NS5 Protein Intrudes in the Cellular Spliceosome and Modulates Splicing
title_full_unstemmed	The Dengue Virus NS5 Protein Intrudes in the Cellular Spliceosome and Modulates Splicing
title_sort	dengue virus ns5 protein intrudes in the cellular spliceosome and modulates splicing
url	http://hdl.handle.net/20.500.12110/paper_15537366_v12_n8_p_DeMaio
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The Dengue Virus NS5 Protein Intrudes in the Cellular Spliceosome and Modulates Splicing

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