Cargando…

Comparative Characterization of the Sindbis Virus Proteome from Mammalian and Invertebrate Hosts Identifies nsP2 as a Component of the Virion and Sorting Nexin 5 as a Significant Host Factor for Alphavirus Replication

Recent advances in mass spectrometry methods and instrumentation now allow for more accurate identification of proteins in low abundance. This technology was applied to Sindbis virus, the prototypical alphavirus, to investigate the viral proteome. To determine if host proteins are specifically packa...

Descripción completa

Detalles Bibliográficos
Autores principales: Schuchman, Ryan, Kilianski, Andy, Piper, Amanda, Vancini, Ricardo, Ribeiro, José M. C., Sprague, Thomas R., Nasar, Farooq, Boyd, Gabrielle, Hernandez, Raquel, Glaros, Trevor
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Microbiology 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6026752/
https://www.ncbi.nlm.nih.gov/pubmed/29743363
http://dx.doi.org/10.1128/JVI.00694-18
_version_ 1783336494978564096
author Schuchman, Ryan
Kilianski, Andy
Piper, Amanda
Vancini, Ricardo
Ribeiro, José M. C.
Sprague, Thomas R.
Nasar, Farooq
Boyd, Gabrielle
Hernandez, Raquel
Glaros, Trevor
author_facet Schuchman, Ryan
Kilianski, Andy
Piper, Amanda
Vancini, Ricardo
Ribeiro, José M. C.
Sprague, Thomas R.
Nasar, Farooq
Boyd, Gabrielle
Hernandez, Raquel
Glaros, Trevor
author_sort Schuchman, Ryan
collection PubMed
description Recent advances in mass spectrometry methods and instrumentation now allow for more accurate identification of proteins in low abundance. This technology was applied to Sindbis virus, the prototypical alphavirus, to investigate the viral proteome. To determine if host proteins are specifically packaged into alphavirus virions, Sindbis virus (SINV) was grown in multiple host cells representing vertebrate and mosquito hosts, and total protein content of purified virions was determined. This analysis identified host factors not previously associated with alphavirus entry, replication, or egress. One host protein, sorting nexin 5 (SNX5), was shown to be critical for the replication of three different alphaviruses, Sindbis, Mayaro, and Chikungunya viruses. The most significant finding was that in addition to the host proteins, SINV nonstructural protein 2 (nsP2) was detected within virions grown in all host cells examined. The protein and RNA-interacting capabilities of nsP2 coupled with its presence in the virion support a role for nsP2 during packaging and/or entry of progeny virus. This function has not been identified for this protein. Taken together, this strategy identified at least one host factor integrally involved in alphavirus replication. Identification of other host proteins provides insight into alphavirus-host interactions during viral replication in both vertebrate and invertebrate hosts. This method of virus proteome analysis may also be useful for the identification of protein candidates for host-based therapeutics. IMPORTANCE Pathogenic alphaviruses, such as Chikungunya and Mayaro viruses, continue to plague public health in developing and developed countries alike. Alphaviruses belong to a group of viruses vectored in nature by hematophagous (blood-feeding) insects and are termed arboviruses (arthropod-borne viruses). This group of viruses contains many human pathogens, such as dengue fever, West Nile, and Yellow fever viruses. With few exceptions, there are no vaccines or prophylactics for these agents, leaving one-third of the world population at risk of infection. Identifying effective antivirals has been a long-term goal for combating these diseases not only because of the lack of vaccines but also because they are effective during an ongoing epidemic. Mass spectrometry-based analysis of the Sindbis virus proteome can be effective in identifying host genes involved in virus replication and novel functions for virus proteins. Identification of these factors is invaluable for the prophylaxis of this group of viruses.
format Online
Article
Text
id pubmed-6026752
institution National Center for Biotechnology Information
language English
publishDate 2018
publisher American Society for Microbiology
record_format MEDLINE/PubMed
spelling pubmed-60267522018-07-06 Comparative Characterization of the Sindbis Virus Proteome from Mammalian and Invertebrate Hosts Identifies nsP2 as a Component of the Virion and Sorting Nexin 5 as a Significant Host Factor for Alphavirus Replication Schuchman, Ryan Kilianski, Andy Piper, Amanda Vancini, Ricardo Ribeiro, José M. C. Sprague, Thomas R. Nasar, Farooq Boyd, Gabrielle Hernandez, Raquel Glaros, Trevor J Virol Virus-Cell Interactions Recent advances in mass spectrometry methods and instrumentation now allow for more accurate identification of proteins in low abundance. This technology was applied to Sindbis virus, the prototypical alphavirus, to investigate the viral proteome. To determine if host proteins are specifically packaged into alphavirus virions, Sindbis virus (SINV) was grown in multiple host cells representing vertebrate and mosquito hosts, and total protein content of purified virions was determined. This analysis identified host factors not previously associated with alphavirus entry, replication, or egress. One host protein, sorting nexin 5 (SNX5), was shown to be critical for the replication of three different alphaviruses, Sindbis, Mayaro, and Chikungunya viruses. The most significant finding was that in addition to the host proteins, SINV nonstructural protein 2 (nsP2) was detected within virions grown in all host cells examined. The protein and RNA-interacting capabilities of nsP2 coupled with its presence in the virion support a role for nsP2 during packaging and/or entry of progeny virus. This function has not been identified for this protein. Taken together, this strategy identified at least one host factor integrally involved in alphavirus replication. Identification of other host proteins provides insight into alphavirus-host interactions during viral replication in both vertebrate and invertebrate hosts. This method of virus proteome analysis may also be useful for the identification of protein candidates for host-based therapeutics. IMPORTANCE Pathogenic alphaviruses, such as Chikungunya and Mayaro viruses, continue to plague public health in developing and developed countries alike. Alphaviruses belong to a group of viruses vectored in nature by hematophagous (blood-feeding) insects and are termed arboviruses (arthropod-borne viruses). This group of viruses contains many human pathogens, such as dengue fever, West Nile, and Yellow fever viruses. With few exceptions, there are no vaccines or prophylactics for these agents, leaving one-third of the world population at risk of infection. Identifying effective antivirals has been a long-term goal for combating these diseases not only because of the lack of vaccines but also because they are effective during an ongoing epidemic. Mass spectrometry-based analysis of the Sindbis virus proteome can be effective in identifying host genes involved in virus replication and novel functions for virus proteins. Identification of these factors is invaluable for the prophylaxis of this group of viruses. American Society for Microbiology 2018-06-29 /pmc/articles/PMC6026752/ /pubmed/29743363 http://dx.doi.org/10.1128/JVI.00694-18 Text en Copyright © 2018 Schuchman et al. https://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Virus-Cell Interactions
Schuchman, Ryan
Kilianski, Andy
Piper, Amanda
Vancini, Ricardo
Ribeiro, José M. C.
Sprague, Thomas R.
Nasar, Farooq
Boyd, Gabrielle
Hernandez, Raquel
Glaros, Trevor
Comparative Characterization of the Sindbis Virus Proteome from Mammalian and Invertebrate Hosts Identifies nsP2 as a Component of the Virion and Sorting Nexin 5 as a Significant Host Factor for Alphavirus Replication
title Comparative Characterization of the Sindbis Virus Proteome from Mammalian and Invertebrate Hosts Identifies nsP2 as a Component of the Virion and Sorting Nexin 5 as a Significant Host Factor for Alphavirus Replication
title_full Comparative Characterization of the Sindbis Virus Proteome from Mammalian and Invertebrate Hosts Identifies nsP2 as a Component of the Virion and Sorting Nexin 5 as a Significant Host Factor for Alphavirus Replication
title_fullStr Comparative Characterization of the Sindbis Virus Proteome from Mammalian and Invertebrate Hosts Identifies nsP2 as a Component of the Virion and Sorting Nexin 5 as a Significant Host Factor for Alphavirus Replication
title_full_unstemmed Comparative Characterization of the Sindbis Virus Proteome from Mammalian and Invertebrate Hosts Identifies nsP2 as a Component of the Virion and Sorting Nexin 5 as a Significant Host Factor for Alphavirus Replication
title_short Comparative Characterization of the Sindbis Virus Proteome from Mammalian and Invertebrate Hosts Identifies nsP2 as a Component of the Virion and Sorting Nexin 5 as a Significant Host Factor for Alphavirus Replication
title_sort comparative characterization of the sindbis virus proteome from mammalian and invertebrate hosts identifies nsp2 as a component of the virion and sorting nexin 5 as a significant host factor for alphavirus replication
topic Virus-Cell Interactions
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6026752/
https://www.ncbi.nlm.nih.gov/pubmed/29743363
http://dx.doi.org/10.1128/JVI.00694-18
work_keys_str_mv AT schuchmanryan comparativecharacterizationofthesindbisvirusproteomefrommammalianandinvertebratehostsidentifiesnsp2asacomponentofthevirionandsortingnexin5asasignificanthostfactorforalphavirusreplication
AT kilianskiandy comparativecharacterizationofthesindbisvirusproteomefrommammalianandinvertebratehostsidentifiesnsp2asacomponentofthevirionandsortingnexin5asasignificanthostfactorforalphavirusreplication
AT piperamanda comparativecharacterizationofthesindbisvirusproteomefrommammalianandinvertebratehostsidentifiesnsp2asacomponentofthevirionandsortingnexin5asasignificanthostfactorforalphavirusreplication
AT vanciniricardo comparativecharacterizationofthesindbisvirusproteomefrommammalianandinvertebratehostsidentifiesnsp2asacomponentofthevirionandsortingnexin5asasignificanthostfactorforalphavirusreplication
AT ribeirojosemc comparativecharacterizationofthesindbisvirusproteomefrommammalianandinvertebratehostsidentifiesnsp2asacomponentofthevirionandsortingnexin5asasignificanthostfactorforalphavirusreplication
AT spraguethomasr comparativecharacterizationofthesindbisvirusproteomefrommammalianandinvertebratehostsidentifiesnsp2asacomponentofthevirionandsortingnexin5asasignificanthostfactorforalphavirusreplication
AT nasarfarooq comparativecharacterizationofthesindbisvirusproteomefrommammalianandinvertebratehostsidentifiesnsp2asacomponentofthevirionandsortingnexin5asasignificanthostfactorforalphavirusreplication
AT boydgabrielle comparativecharacterizationofthesindbisvirusproteomefrommammalianandinvertebratehostsidentifiesnsp2asacomponentofthevirionandsortingnexin5asasignificanthostfactorforalphavirusreplication
AT hernandezraquel comparativecharacterizationofthesindbisvirusproteomefrommammalianandinvertebratehostsidentifiesnsp2asacomponentofthevirionandsortingnexin5asasignificanthostfactorforalphavirusreplication
AT glarostrevor comparativecharacterizationofthesindbisvirusproteomefrommammalianandinvertebratehostsidentifiesnsp2asacomponentofthevirionandsortingnexin5asasignificanthostfactorforalphavirusreplication