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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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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