A Comprehensive Screening of the Interactors of Areca Palm Necrotic Ringspot Virus (ANRSV) HCPro2 Highlights the Proviral Roles of eIF4A and PGK in Viral Infection

The areca palm (<i>Areca catechu</i> L.), a medicinal tropical crop, hosts three novel viruses, areca palm necrotic ringspot virus (ANRSV), areca palm necrotic spindle-spot virus (ANSSV), and ANRSV2, which form a new genus <i>Arepavirus</i> in the family <i>Potyviridae&...

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Main Authors: Li Qin, Peilan Liu, Wentao Shen, Zhaoji Dai, Hongguang Cui
Format: Article
Language:English
Published: MDPI AG 2025-05-01
Series:Plants
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Online Access:https://www.mdpi.com/2223-7747/14/11/1673
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author Li Qin
Peilan Liu
Wentao Shen
Zhaoji Dai
Hongguang Cui
author_facet Li Qin
Peilan Liu
Wentao Shen
Zhaoji Dai
Hongguang Cui
author_sort Li Qin
collection DOAJ
description The areca palm (<i>Areca catechu</i> L.), a medicinal tropical crop, hosts three novel viruses, areca palm necrotic ringspot virus (ANRSV), areca palm necrotic spindle-spot virus (ANSSV), and ANRSV2, which form a new genus <i>Arepavirus</i> in the family <i>Potyviridae</i>. Both viruses feature a unique tandem leader protease arrangement (HCPro1-HCPro2). To elucidate HCPro2’s role, this study identified its interaction partners in infected cells using affinity purification coupled with liquid chromatography-tandem mass spectrometry, a yeast two-hybrid system, and co-immunoprecipitation. Thirteen host proteins and five viral factors (HCPro1, 6K2, VPg, NIa-Pro, NIb) were validated as HCPro2 interactors. Among the host proteins interacting with HCPro2, the expression of five genes (<i>NbeIF4A</i>, <i>NbSAMS1α</i>, <i>NbTEF1α</i>, <i>NbUEP1,</i> and <i>NbRan2</i>) was upregulated under the condition of viral infection, while the expression of another five genes (<i>NbpsbS1</i>, <i>NbPGK</i>, <i>NbchIP</i>, <i>NbClpC1A</i>, and <i>NbCysPrx</i>) was downregulated. Functional assays showed that silencing <i>NbeIF4A</i> or <i>NbPGK</i> significantly reduced viral accumulation <i>in Nicotiana benthamiana</i>. These findings reveal HCPro2’s network of virus-host interaction, highlighting its critical role in viral pathogenesis. Further exploration of these interactions may clarify the evolutionary significance of tandem leader proteases and inform novel plant antiviral strategies.
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issn 2223-7747
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spelling doaj-art-b36ff57fc591462fbe784edea26000ad2025-08-20T03:46:45ZengMDPI AGPlants2223-77472025-05-011411167310.3390/plants14111673A Comprehensive Screening of the Interactors of Areca Palm Necrotic Ringspot Virus (ANRSV) HCPro2 Highlights the Proviral Roles of eIF4A and PGK in Viral InfectionLi Qin0Peilan Liu1Wentao Shen2Zhaoji Dai3Hongguang Cui4Key Laboratory of Green Prevention and Control of Tropical Plant Diseases and Pests (Ministry of Education), School of Tropical Agriculture and Forestry, Hainan University, Haikou 570228, ChinaKey Laboratory of Green Prevention and Control of Tropical Plant Diseases and Pests (Ministry of Education), School of Tropical Agriculture and Forestry, Hainan University, Haikou 570228, ChinaInstitute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, Haikou 571101, ChinaKey Laboratory of Green Prevention and Control of Tropical Plant Diseases and Pests (Ministry of Education), School of Tropical Agriculture and Forestry, Hainan University, Haikou 570228, ChinaKey Laboratory of Green Prevention and Control of Tropical Plant Diseases and Pests (Ministry of Education), School of Tropical Agriculture and Forestry, Hainan University, Haikou 570228, ChinaThe areca palm (<i>Areca catechu</i> L.), a medicinal tropical crop, hosts three novel viruses, areca palm necrotic ringspot virus (ANRSV), areca palm necrotic spindle-spot virus (ANSSV), and ANRSV2, which form a new genus <i>Arepavirus</i> in the family <i>Potyviridae</i>. Both viruses feature a unique tandem leader protease arrangement (HCPro1-HCPro2). To elucidate HCPro2’s role, this study identified its interaction partners in infected cells using affinity purification coupled with liquid chromatography-tandem mass spectrometry, a yeast two-hybrid system, and co-immunoprecipitation. Thirteen host proteins and five viral factors (HCPro1, 6K2, VPg, NIa-Pro, NIb) were validated as HCPro2 interactors. Among the host proteins interacting with HCPro2, the expression of five genes (<i>NbeIF4A</i>, <i>NbSAMS1α</i>, <i>NbTEF1α</i>, <i>NbUEP1,</i> and <i>NbRan2</i>) was upregulated under the condition of viral infection, while the expression of another five genes (<i>NbpsbS1</i>, <i>NbPGK</i>, <i>NbchIP</i>, <i>NbClpC1A</i>, and <i>NbCysPrx</i>) was downregulated. Functional assays showed that silencing <i>NbeIF4A</i> or <i>NbPGK</i> significantly reduced viral accumulation <i>in Nicotiana benthamiana</i>. These findings reveal HCPro2’s network of virus-host interaction, highlighting its critical role in viral pathogenesis. Further exploration of these interactions may clarify the evolutionary significance of tandem leader proteases and inform novel plant antiviral strategies.https://www.mdpi.com/2223-7747/14/11/1673<i>Potyviridae</i>leader proteasecysteine proteaseeukaryotic initiation factor 4Aphosphoglycerate kinase
spellingShingle Li Qin
Peilan Liu
Wentao Shen
Zhaoji Dai
Hongguang Cui
A Comprehensive Screening of the Interactors of Areca Palm Necrotic Ringspot Virus (ANRSV) HCPro2 Highlights the Proviral Roles of eIF4A and PGK in Viral Infection
Plants
<i>Potyviridae</i>
leader protease
cysteine protease
eukaryotic initiation factor 4A
phosphoglycerate kinase
title A Comprehensive Screening of the Interactors of Areca Palm Necrotic Ringspot Virus (ANRSV) HCPro2 Highlights the Proviral Roles of eIF4A and PGK in Viral Infection
title_full A Comprehensive Screening of the Interactors of Areca Palm Necrotic Ringspot Virus (ANRSV) HCPro2 Highlights the Proviral Roles of eIF4A and PGK in Viral Infection
title_fullStr A Comprehensive Screening of the Interactors of Areca Palm Necrotic Ringspot Virus (ANRSV) HCPro2 Highlights the Proviral Roles of eIF4A and PGK in Viral Infection
title_full_unstemmed A Comprehensive Screening of the Interactors of Areca Palm Necrotic Ringspot Virus (ANRSV) HCPro2 Highlights the Proviral Roles of eIF4A and PGK in Viral Infection
title_short A Comprehensive Screening of the Interactors of Areca Palm Necrotic Ringspot Virus (ANRSV) HCPro2 Highlights the Proviral Roles of eIF4A and PGK in Viral Infection
title_sort comprehensive screening of the interactors of areca palm necrotic ringspot virus anrsv hcpro2 highlights the proviral roles of eif4a and pgk in viral infection
topic <i>Potyviridae</i>
leader protease
cysteine protease
eukaryotic initiation factor 4A
phosphoglycerate kinase
url https://www.mdpi.com/2223-7747/14/11/1673
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