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VPg Impact on Ryegrass Mottle Virus Serine-like 3C Protease Proteolysis and Structure

Sobemoviruses encode serine-like 3C proteases (Pro) that participate in the processing and maturation of other virus-encoded proteins. Its cis and trans activity is mediated by the naturally unfolded virus-genome-linked protein (VPg). Nuclear magnetic resonance studies show a Pro–VPg complex interac...

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Autores principales: Kalnins, Gints, Ludviga, Rebeka, Kalnciema, Ieva, Resevica, Gunta, Zeltina, Vilija, Bogans, Janis, Tars, Kaspars, Zeltins, Andris, Balke, Ina
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10048871/
https://www.ncbi.nlm.nih.gov/pubmed/36982419
http://dx.doi.org/10.3390/ijms24065347
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author Kalnins, Gints
Ludviga, Rebeka
Kalnciema, Ieva
Resevica, Gunta
Zeltina, Vilija
Bogans, Janis
Tars, Kaspars
Zeltins, Andris
Balke, Ina
author_facet Kalnins, Gints
Ludviga, Rebeka
Kalnciema, Ieva
Resevica, Gunta
Zeltina, Vilija
Bogans, Janis
Tars, Kaspars
Zeltins, Andris
Balke, Ina
author_sort Kalnins, Gints
collection PubMed
description Sobemoviruses encode serine-like 3C proteases (Pro) that participate in the processing and maturation of other virus-encoded proteins. Its cis and trans activity is mediated by the naturally unfolded virus-genome-linked protein (VPg). Nuclear magnetic resonance studies show a Pro–VPg complex interaction and VPg tertiary structure; however, information regarding structural changes of the Pro–VPg complex during interaction is lacking. Here, we solved a full Pro–VPg 3D structure of ryegrass mottle virus (RGMoV) that demonstrates the structural changes in three different conformations due to VPg interaction with Pro. We identified a unique site of VPg interaction with Pro that was not observed in other sobemoviruses, and observed different conformations of the Pro β2 barrel. This is the first report of a full plant Pro crystal structure with its VPg cofactor. We also confirmed the existence of an unusual previously unmapped cleavage site for sobemovirus Pro in the transmembrane domain: E/A. We demonstrated that RGMoV Pro in cis activity is not regulated by VPg and that in trans, VPg can also mediate Pro in free form. Additionally, we observed Ca(2+) and Zn(2+) inhibitory effects on the Pro cleavage activity.
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spelling pubmed-100488712023-03-29 VPg Impact on Ryegrass Mottle Virus Serine-like 3C Protease Proteolysis and Structure Kalnins, Gints Ludviga, Rebeka Kalnciema, Ieva Resevica, Gunta Zeltina, Vilija Bogans, Janis Tars, Kaspars Zeltins, Andris Balke, Ina Int J Mol Sci Article Sobemoviruses encode serine-like 3C proteases (Pro) that participate in the processing and maturation of other virus-encoded proteins. Its cis and trans activity is mediated by the naturally unfolded virus-genome-linked protein (VPg). Nuclear magnetic resonance studies show a Pro–VPg complex interaction and VPg tertiary structure; however, information regarding structural changes of the Pro–VPg complex during interaction is lacking. Here, we solved a full Pro–VPg 3D structure of ryegrass mottle virus (RGMoV) that demonstrates the structural changes in three different conformations due to VPg interaction with Pro. We identified a unique site of VPg interaction with Pro that was not observed in other sobemoviruses, and observed different conformations of the Pro β2 barrel. This is the first report of a full plant Pro crystal structure with its VPg cofactor. We also confirmed the existence of an unusual previously unmapped cleavage site for sobemovirus Pro in the transmembrane domain: E/A. We demonstrated that RGMoV Pro in cis activity is not regulated by VPg and that in trans, VPg can also mediate Pro in free form. Additionally, we observed Ca(2+) and Zn(2+) inhibitory effects on the Pro cleavage activity. MDPI 2023-03-10 /pmc/articles/PMC10048871/ /pubmed/36982419 http://dx.doi.org/10.3390/ijms24065347 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Kalnins, Gints
Ludviga, Rebeka
Kalnciema, Ieva
Resevica, Gunta
Zeltina, Vilija
Bogans, Janis
Tars, Kaspars
Zeltins, Andris
Balke, Ina
VPg Impact on Ryegrass Mottle Virus Serine-like 3C Protease Proteolysis and Structure
title VPg Impact on Ryegrass Mottle Virus Serine-like 3C Protease Proteolysis and Structure
title_full VPg Impact on Ryegrass Mottle Virus Serine-like 3C Protease Proteolysis and Structure
title_fullStr VPg Impact on Ryegrass Mottle Virus Serine-like 3C Protease Proteolysis and Structure
title_full_unstemmed VPg Impact on Ryegrass Mottle Virus Serine-like 3C Protease Proteolysis and Structure
title_short VPg Impact on Ryegrass Mottle Virus Serine-like 3C Protease Proteolysis and Structure
title_sort vpg impact on ryegrass mottle virus serine-like 3c protease proteolysis and structure
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10048871/
https://www.ncbi.nlm.nih.gov/pubmed/36982419
http://dx.doi.org/10.3390/ijms24065347
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