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

PLoS Pathogens

Berta Pozzi¹  Anabella Srebrow¹  Nevan Krogan²  Priya Shah²  Federico A. De Maio³  Raul Andino³  Nestor G. Iglesias³  Marcelo J. Yanovsky³  Pablo Mammi³  Leopoldo G. Gebhard⁴  Andrea V. Gamarnik⁴  Guillermo Risso⁴  Estefania Mancini⁴ 

[1] Department of Cellular and Molecular Pharmacology, University of California, San Francisco, California, United States of America;Department of Microbiology and Immunology, University of California, San Francisco, San Francisco, California, United States of America;Fundación Instituto Leloir-CONICET, Buenos Aires, Argentina;Instituto de Fisiología, Biología Molecular y Neurociencias (IFIBYNE, UBA-CONICET), Departamento de Fisiología, Biología Molecular y Celular, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires

关键词: Alternative splicing;    Viral replication;    Introns;    Small interfering RNAs;    Viral transmission;    infection;    RNA splicing;    Dengue virus;    Spliceosomes;   

DOI  :  10.1371/journal.ppat.1005841

学科分类：生物科学（综合）

来源: Public Library of Science

【 摘 要 】

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.

【 授权许可】

CC BY