dsRNA Binding Domain of PKR Is Proteolytically Released by Enterovirus A71 to Facilitate Viral Replication

Enterovirus 71 (EV-A71) causes hand, foot and mouth disease in young children and infants, but can also cause severe neurological complications or even death. The double-stranded RNA (dsRNA)-dependent protein kinase R (PKR), an interferon-induced antiviral protein, phosphorylates the regulatory α-su...

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Main Authors: Yu-Hsiu Chang, Kean Seng Lau, Rei-Lin Kuo, Jim-Tong Horng
Format: Article
Language:English
Published: Frontiers Media S.A. 2017-06-01
Series:Frontiers in Cellular and Infection Microbiology
Subjects:
PKR
Online Access:http://journal.frontiersin.org/article/10.3389/fcimb.2017.00284/full
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spelling doaj-96243466e5244c37a2880ed3bbf20ecb2020-11-24T21:02:55ZengFrontiers Media S.A.Frontiers in Cellular and Infection Microbiology2235-29882017-06-01710.3389/fcimb.2017.00284267686dsRNA Binding Domain of PKR Is Proteolytically Released by Enterovirus A71 to Facilitate Viral ReplicationYu-Hsiu Chang0Yu-Hsiu Chang1Kean Seng Lau2Rei-Lin Kuo3Rei-Lin Kuo4Jim-Tong Horng5Jim-Tong Horng6Jim-Tong Horng7Jim-Tong Horng8Jim-Tong Horng9Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung UniversityTaoyuan, TaiwanNational Defense Medical Center, Institute of Preventive MedicineTaipei, TaiwanDepartment of Biochemistry and Molecular Biology, College of Medicine, Chang Gung UniversityTaoyuan, TaiwanResearch Center for Emerging Viral Infections, College of Medicine, Chang Gung UniversityTaoyuan, TaiwanMolecular Infectious Disease Research Center, Chang Gung Memorial HospitalTaoyuan, TaiwanGraduate Institute of Biomedical Sciences, College of Medicine, Chang Gung UniversityTaoyuan, TaiwanDepartment of Biochemistry and Molecular Biology, College of Medicine, Chang Gung UniversityTaoyuan, TaiwanResearch Center for Emerging Viral Infections, College of Medicine, Chang Gung UniversityTaoyuan, TaiwanMolecular Infectious Disease Research Center, Chang Gung Memorial HospitalTaoyuan, TaiwanResearch Center for Chinese Herbal Medicine and Research Center for Food and Cosmetic Safety, College of Human Ecology, Chang Gung University of Science and TechnologyTaoyuan, TaiwanEnterovirus 71 (EV-A71) causes hand, foot and mouth disease in young children and infants, but can also cause severe neurological complications or even death. The double-stranded RNA (dsRNA)-dependent protein kinase R (PKR), an interferon-induced antiviral protein, phosphorylates the regulatory α-subunit of the eukaryotic translation initiation factor 2 in response to viral infection, thereby blocking the translation of cellular and viral mRNA and promoting apoptosis. The cleavage of PKR after infection with poliovirus, a prototype enterovirus, has been reported by others, but the underlying mechanism of this cleavage and its role in viral replication remain unclear. In the present study, we show that viral 3C protease cleaves PKR at a site, Q188, which differs from the site cleaved during apoptosis, D251. In contrast to the conventional phosphorylation of PKR by dsRNA, EV-A71 3C physically interacts with PKR to mediate the phosphorylation of PKR; this effect is dependent on 3C protease activity. Overexpression of a catalytically inactive PKR mutant (K296H) accelerates viral protein accumulation and increases virus titer, whereas a K64E substitution in the dsRNA binding site abolishes this advantage. We also demonstrate that PKR cleavage mediated by EV-A71 3C protease produces a short N-terminal PKR fragment that can enhance EV-A71 replication, in terms of viral RNA, viral protein, and viral titers. We conclude that PKR is co-opted by EV-A71 via viral protease 3C-mediated proteolytic activation to facilitate viral replication.http://journal.frontiersin.org/article/10.3389/fcimb.2017.00284/fullEV-A713C proteasePKREnterovirusinteraction effects
collection DOAJ
language English
format Article
sources DOAJ
author Yu-Hsiu Chang
Yu-Hsiu Chang
Kean Seng Lau
Rei-Lin Kuo
Rei-Lin Kuo
Jim-Tong Horng
Jim-Tong Horng
Jim-Tong Horng
Jim-Tong Horng
Jim-Tong Horng
spellingShingle Yu-Hsiu Chang
Yu-Hsiu Chang
Kean Seng Lau
Rei-Lin Kuo
Rei-Lin Kuo
Jim-Tong Horng
Jim-Tong Horng
Jim-Tong Horng
Jim-Tong Horng
Jim-Tong Horng
dsRNA Binding Domain of PKR Is Proteolytically Released by Enterovirus A71 to Facilitate Viral Replication
Frontiers in Cellular and Infection Microbiology
EV-A71
3C protease
PKR
Enterovirus
interaction effects
author_facet Yu-Hsiu Chang
Yu-Hsiu Chang
Kean Seng Lau
Rei-Lin Kuo
Rei-Lin Kuo
Jim-Tong Horng
Jim-Tong Horng
Jim-Tong Horng
Jim-Tong Horng
Jim-Tong Horng
author_sort Yu-Hsiu Chang
title dsRNA Binding Domain of PKR Is Proteolytically Released by Enterovirus A71 to Facilitate Viral Replication
title_short dsRNA Binding Domain of PKR Is Proteolytically Released by Enterovirus A71 to Facilitate Viral Replication
title_full dsRNA Binding Domain of PKR Is Proteolytically Released by Enterovirus A71 to Facilitate Viral Replication
title_fullStr dsRNA Binding Domain of PKR Is Proteolytically Released by Enterovirus A71 to Facilitate Viral Replication
title_full_unstemmed dsRNA Binding Domain of PKR Is Proteolytically Released by Enterovirus A71 to Facilitate Viral Replication
title_sort dsrna binding domain of pkr is proteolytically released by enterovirus a71 to facilitate viral replication
publisher Frontiers Media S.A.
series Frontiers in Cellular and Infection Microbiology
issn 2235-2988
publishDate 2017-06-01
description Enterovirus 71 (EV-A71) causes hand, foot and mouth disease in young children and infants, but can also cause severe neurological complications or even death. The double-stranded RNA (dsRNA)-dependent protein kinase R (PKR), an interferon-induced antiviral protein, phosphorylates the regulatory α-subunit of the eukaryotic translation initiation factor 2 in response to viral infection, thereby blocking the translation of cellular and viral mRNA and promoting apoptosis. The cleavage of PKR after infection with poliovirus, a prototype enterovirus, has been reported by others, but the underlying mechanism of this cleavage and its role in viral replication remain unclear. In the present study, we show that viral 3C protease cleaves PKR at a site, Q188, which differs from the site cleaved during apoptosis, D251. In contrast to the conventional phosphorylation of PKR by dsRNA, EV-A71 3C physically interacts with PKR to mediate the phosphorylation of PKR; this effect is dependent on 3C protease activity. Overexpression of a catalytically inactive PKR mutant (K296H) accelerates viral protein accumulation and increases virus titer, whereas a K64E substitution in the dsRNA binding site abolishes this advantage. We also demonstrate that PKR cleavage mediated by EV-A71 3C protease produces a short N-terminal PKR fragment that can enhance EV-A71 replication, in terms of viral RNA, viral protein, and viral titers. We conclude that PKR is co-opted by EV-A71 via viral protease 3C-mediated proteolytic activation to facilitate viral replication.
topic EV-A71
3C protease
PKR
Enterovirus
interaction effects
url http://journal.frontiersin.org/article/10.3389/fcimb.2017.00284/full
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