An Evolutionarily Conserved Pathway Essential for Orsay Virus Infection of <italic toggle="yes">Caenorhabditis elegans</italic>

ABSTRACT Many fundamental biological discoveries have been made in Caenorhabditis elegans. The discovery of Orsay virus has enabled studies of host-virus interactions in this model organism. To identify host factors critical for Orsay virus infection, we designed a forward genetic screen that utiliz...

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Autores principales:	Hongbing Jiang, Kevin Chen, Luis E. Sandoval, Christian Leung, David Wang
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2017
Materias:	Caenorhabditis elegans Orsay virus TNK2 WASP sid-3 viro-2 Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/ddfc5784f66d4c86a08ea51575dad1e6
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spelling	oai:doaj.org-article:ddfc5784f66d4c86a08ea51575dad1e62021-11-15T15:51:51ZAn Evolutionarily Conserved Pathway Essential for Orsay Virus Infection of <italic toggle="yes">Caenorhabditis elegans</italic>10.1128/mBio.00940-172150-7511https://doaj.org/article/ddfc5784f66d4c86a08ea51575dad1e62017-11-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00940-17https://doaj.org/toc/2150-7511ABSTRACT Many fundamental biological discoveries have been made in Caenorhabditis elegans. The discovery of Orsay virus has enabled studies of host-virus interactions in this model organism. To identify host factors critical for Orsay virus infection, we designed a forward genetic screen that utilizes a virally induced green fluorescent protein (GFP) reporter. Following chemical mutagenesis, two Viro (virus induced reporter off) mutants that failed to express GFP were mapped to sid-3, a nonreceptor tyrosine kinase, and B0280.13 (renamed viro-2), an ortholog of human Wiskott-Aldrich syndrome protein (WASP). Both mutants yielded Orsay virus RNA levels comparable to that of the residual input virus, suggesting that they are not permissive for Orsay virus replication. In addition, we demonstrated that both genes affect an early prereplication stage of Orsay virus infection. Furthermore, it is known that the human ortholog of SID-3, activated CDC42-associated kinase (ACK1/TNK2), is capable of phosphorylating human WASP, suggesting that VIRO-2 may be a substrate for SID-3 in C. elegans. A targeted RNA interference (RNAi) knockdown screen further identified the C. elegans gene nck-1, which has a human ortholog that interacts with TNK2 and WASP, as required for Orsay virus infection. Thus, genetic screening in C. elegans identified critical roles in virus infection for evolutionarily conserved genes in a known human pathway. IMPORTANCE Orsay virus is the only known virus capable of naturally infecting the model organism Caenorhabditis elegans, which shares many evolutionarily conserved genes with humans. We exploited the robust genetic tractability of C. elegans to identify three host genes, sid-3, viro-2, and nck-1, which are essential for Orsay virus infection. Mutant animals that lack these three genes are highly defective in viral replication. Strikingly, the human orthologs of these three genes, activated CDC42-associated kinase (TNK2), Wiskott-Aldrich syndrome protein (WASP), and noncatalytic region of tyrosine kinase adaptor protein 1 (NCK1) are part of a known signaling pathway in mammals. These results suggest that TNK2, WASP, and NCK1 may play important roles in mammalian virus infection.Hongbing JiangKevin ChenLuis E. SandovalChristian LeungDavid WangAmerican Society for MicrobiologyarticleCaenorhabditis elegansOrsay virusTNK2WASPsid-3viro-2MicrobiologyQR1-502ENmBio, Vol 8, Iss 5 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Caenorhabditis elegans Orsay virus TNK2 WASP sid-3 viro-2 Microbiology QR1-502
spellingShingle	Caenorhabditis elegans Orsay virus TNK2 WASP sid-3 viro-2 Microbiology QR1-502 Hongbing Jiang Kevin Chen Luis E. Sandoval Christian Leung David Wang An Evolutionarily Conserved Pathway Essential for Orsay Virus Infection of <italic toggle="yes">Caenorhabditis elegans</italic>
description	ABSTRACT Many fundamental biological discoveries have been made in Caenorhabditis elegans. The discovery of Orsay virus has enabled studies of host-virus interactions in this model organism. To identify host factors critical for Orsay virus infection, we designed a forward genetic screen that utilizes a virally induced green fluorescent protein (GFP) reporter. Following chemical mutagenesis, two Viro (virus induced reporter off) mutants that failed to express GFP were mapped to sid-3, a nonreceptor tyrosine kinase, and B0280.13 (renamed viro-2), an ortholog of human Wiskott-Aldrich syndrome protein (WASP). Both mutants yielded Orsay virus RNA levels comparable to that of the residual input virus, suggesting that they are not permissive for Orsay virus replication. In addition, we demonstrated that both genes affect an early prereplication stage of Orsay virus infection. Furthermore, it is known that the human ortholog of SID-3, activated CDC42-associated kinase (ACK1/TNK2), is capable of phosphorylating human WASP, suggesting that VIRO-2 may be a substrate for SID-3 in C. elegans. A targeted RNA interference (RNAi) knockdown screen further identified the C. elegans gene nck-1, which has a human ortholog that interacts with TNK2 and WASP, as required for Orsay virus infection. Thus, genetic screening in C. elegans identified critical roles in virus infection for evolutionarily conserved genes in a known human pathway. IMPORTANCE Orsay virus is the only known virus capable of naturally infecting the model organism Caenorhabditis elegans, which shares many evolutionarily conserved genes with humans. We exploited the robust genetic tractability of C. elegans to identify three host genes, sid-3, viro-2, and nck-1, which are essential for Orsay virus infection. Mutant animals that lack these three genes are highly defective in viral replication. Strikingly, the human orthologs of these three genes, activated CDC42-associated kinase (TNK2), Wiskott-Aldrich syndrome protein (WASP), and noncatalytic region of tyrosine kinase adaptor protein 1 (NCK1) are part of a known signaling pathway in mammals. These results suggest that TNK2, WASP, and NCK1 may play important roles in mammalian virus infection.
format	article
author	Hongbing Jiang Kevin Chen Luis E. Sandoval Christian Leung David Wang
author_facet	Hongbing Jiang Kevin Chen Luis E. Sandoval Christian Leung David Wang
author_sort	Hongbing Jiang
title	An Evolutionarily Conserved Pathway Essential for Orsay Virus Infection of <italic toggle="yes">Caenorhabditis elegans</italic>
title_short	An Evolutionarily Conserved Pathway Essential for Orsay Virus Infection of <italic toggle="yes">Caenorhabditis elegans</italic>
title_full	An Evolutionarily Conserved Pathway Essential for Orsay Virus Infection of <italic toggle="yes">Caenorhabditis elegans</italic>
title_fullStr	An Evolutionarily Conserved Pathway Essential for Orsay Virus Infection of <italic toggle="yes">Caenorhabditis elegans</italic>
title_full_unstemmed	An Evolutionarily Conserved Pathway Essential for Orsay Virus Infection of <italic toggle="yes">Caenorhabditis elegans</italic>
title_sort	evolutionarily conserved pathway essential for orsay virus infection of <italic toggle="yes">caenorhabditis elegans</italic>
publisher	American Society for Microbiology
publishDate	2017
url	https://doaj.org/article/ddfc5784f66d4c86a08ea51575dad1e6
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An Evolutionarily Conserved Pathway Essential for Orsay Virus Infection of <italic toggle="yes">Caenorhabditis elegans</italic>

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