Cytoplasmic viral RNA-dependent RNA polymerase disrupts the intracellular splicing machinery by entering the nucleus and interfering with Prp8

Yen-Chin Liu; Rei-Lin Kuo; Jing-Yi Lin; Peng-Nien Huang; Yi Huang; Hsuan Liu; Jamine J Arnold; Shu-Jen Chen; Robert Yung-Liang Wang; Craig E Cameron; Shin-Ru Shih

doi:10.1371/journal.ppat.1004199

Cytoplasmic viral RNA-dependent RNA polymerase disrupts the intracellular splicing machinery by entering the nucleus and interfering with Prp8

PLoS Pathog. 2014 Jun 26;10(6):e1004199. doi: 10.1371/journal.ppat.1004199. eCollection 2014 Jun.

Authors

Yen-Chin Liu¹, Rei-Lin Kuo², Jing-Yi Lin³, Peng-Nien Huang⁴, Yi Huang⁵, Hsuan Liu⁵, Jamine J Arnold⁶, Shu-Jen Chen⁵, Robert Yung-Liang Wang⁷, Craig E Cameron⁶, Shin-Ru Shih⁸

Affiliations

¹ Research Center for Emerging Viral Infections, College of Medicine, Chang Gung University, Tao-Yuan, Taiwan; Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Tao-Yuan, Taiwan.
² Research Center for Emerging Viral Infections, College of Medicine, Chang Gung University, Tao-Yuan, Taiwan; Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Tao-Yuan, Taiwan; Department of Medical Biotechnology and Laboratory Science, College of Medicine, Chang Gung University, Tao-Yuan, Taiwan.
³ School of Medical Laboratory Science and Biotechnology, Taipei Medical University, Taipei, Taiwan.
⁴ Research Center for Emerging Viral Infections, College of Medicine, Chang Gung University, Tao-Yuan, Taiwan.
⁵ Molecular Medicine Research Center, Chang Gung University, Tao-Yuan, Taiwan.
⁶ Department of Biochemistry and Molecular Biology, Pennsylvania State University, University Park, Pennsylvania, United States of America.
⁷ Research Center for Emerging Viral Infections, College of Medicine, Chang Gung University, Tao-Yuan, Taiwan; Department of Biomedical Sciences and Graduate Institutes of Biomedical Sciences, College of Medicine, Chang Gung University, Tao-Yuan, Taiwan.
⁸ Research Center for Emerging Viral Infections, College of Medicine, Chang Gung University, Tao-Yuan, Taiwan; Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Tao-Yuan, Taiwan; Department of Medical Biotechnology and Laboratory Science, College of Medicine, Chang Gung University, Tao-Yuan, Taiwan; Clinical Virology Laboratory, Chang Gung Memorial Hospital, Tao-Yuan, Taiwan.

Abstract

The primary role of cytoplasmic viral RNA-dependent RNA polymerase (RdRp) is viral genome replication in the cellular cytoplasm. However, picornaviral RdRp denoted 3D polymerase (3D(pol)) also enters the host nucleus, where its function remains unclear. In this study, we describe a novel mechanism of viral attack in which 3D(pol) enters the nucleus through the nuclear localization signal (NLS) and targets the pre-mRNA processing factor 8 (Prp8) to block pre-mRNA splicing and mRNA synthesis. The fingers domain of 3D(pol) associates with the C-terminal region of Prp8, which contains the Jab1/MPN domain, and interferes in the second catalytic step, resulting in the accumulation of the lariat form of the splicing intermediate. Endogenous pre-mRNAs trapped by the Prp8-3D(pol) complex in enterovirus-infected cells were identified and classed into groups associated with cell growth, proliferation, and differentiation. Our results suggest that picornaviral RdRp disrupts pre-mRNA splicing processes, that differs from viral protease shutting off cellular transcription and translation which contributes to the pathogenesis of viral infection.

Publication types

Comparative Study
Research Support, Non-U.S. Gov't

MeSH terms

Cell Line
Cell Nucleus / metabolism*
Cell Nucleus / virology
Cytoplasm / enzymology*
Cytoplasm / metabolism
Cytoplasm / virology
Enterovirus / metabolism
Enterovirus / pathogenicity
Enterovirus A, Human / metabolism*
Enterovirus A, Human / pathogenicity
Humans
Nuclear Localization Signals
Poliovirus / metabolism
Poliovirus / pathogenicity
Protein Biosynthesis
Protein Interaction Domains and Motifs
RNA Precursors / antagonists & inhibitors
RNA Precursors / metabolism
RNA Splicing*
RNA, Messenger / antagonists & inhibitors
RNA, Messenger / metabolism
RNA-Binding Proteins / antagonists & inhibitors*
RNA-Binding Proteins / chemistry
RNA-Binding Proteins / metabolism
RNA-Directed DNA Polymerase / chemistry
RNA-Directed DNA Polymerase / genetics
RNA-Directed DNA Polymerase / metabolism*
Recombinant Proteins / chemistry
Recombinant Proteins / metabolism
Rhinovirus / metabolism
Rhinovirus / pathogenicity
Species Specificity
Transcription, Genetic
Viral Load
Viral Proteins / chemistry
Viral Proteins / genetics
Viral Proteins / metabolism*
Virulence

Substances

Nuclear Localization Signals
PRPF8 protein, human
RNA Precursors
RNA, Messenger
RNA-Binding Proteins
Recombinant Proteins
Viral Proteins
RNA-Directed DNA Polymerase

Grants and funding

This work was supported by NRPGM (NSC 102-2325-B-182-015). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.