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Cellular proteins associated with the interior and exterior of vesicular stomatitis virus virions.

Moerdyk-Schauwecker M, Hwang SI, Grdzelishvili VZ - PLoS ONE (2014)

Bottom Line: Most of these proteins have not been previously shown to be associated with VSV.Functional enrichment analysis indicated the most overrepresented categories were proteins associated with vesicles, vesicle-mediated transport and protein localization.Our study provides a valuable inventory of virion-associated host proteins for further investigation of their roles in the replication cycle, pathogenesis and immunoreactivity of VSV.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Sciences, University of North Carolina at Charlotte, Charlotte, North Carolina, United States of America; Center for Biomedical Engineering and Science, University of North Carolina at Charlotte, Charlotte, North Carolina, United States of America.

ABSTRACT
Virus particles (virions) often contain not only virus-encoded but also host-encoded proteins. Some of these host proteins are enclosed within the virion structure, while others, in the case of enveloped viruses, are embedded in the host-derived membrane. While many of these host protein incorporations are likely accidental, some may play a role in virus infectivity, replication and/or immunoreactivity in the next host. Host protein incorporations may be especially important in therapeutic applications where large numbers of virus particles are administered. Vesicular stomatitis virus (VSV) is the prototypic rhabdovirus and a candidate vaccine, gene therapy and oncolytic vector. Using mass spectrometry, we previously examined cell type dependent host protein content of VSV virions using intact ("whole") virions purified from three cell lines originating from different species. Here we aimed to determine the localization of host proteins within the VSV virions by analyzing: i) whole VSV virions; and ii) whole VSV virions treated with Proteinase K to remove all proteins outside the viral envelope. A total of 257 proteins were identified, with 181 identified in whole virions and 183 identified in Proteinase K treated virions. Most of these proteins have not been previously shown to be associated with VSV. Functional enrichment analysis indicated the most overrepresented categories were proteins associated with vesicles, vesicle-mediated transport and protein localization. Using western blotting, the presence of several host proteins, including some not previously shown in association with VSV (such as Yes1, Prl1 and Ddx3y), was confirmed and their relative quantities in various virion fractions determined. Our study provides a valuable inventory of virion-associated host proteins for further investigation of their roles in the replication cycle, pathogenesis and immunoreactivity of VSV.

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Related in: MedlinePlus

Functional enrichment analysis.Enrichment analysis and clustering based on gene ontology terms was conducted for (a) all proteins identified and (b) proteins identified in the proteinase K (ProK) treated virions, as described in the materials in methods. The five clusters in each database with the highest enrichment score are depicted here. Numbers above the bars indicate the number of identified proteins associated with each cluster.
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pone-0104688-g006: Functional enrichment analysis.Enrichment analysis and clustering based on gene ontology terms was conducted for (a) all proteins identified and (b) proteins identified in the proteinase K (ProK) treated virions, as described in the materials in methods. The five clusters in each database with the highest enrichment score are depicted here. Numbers above the bars indicate the number of identified proteins associated with each cluster.

Mentions: To obtain an overview of the types of proteins most commonly associated with purified VSV virions, an enrichment analysis was conducted using gene ontology terms. This analysis was done for all three GO databases (biological process, cellular component and molecular function), first using all the host proteins found in our analysis then looking specifically at proteins associated with ProK treated virions (Fig. 6), as this is the highest purity sample and excludes proteins found exclusively on the exterior of the virion. Related enriched terms were then clustered together to give an overall enrichment score. The most highly enriched clusters, when looking at all the proteins, were vesicles and vesicle mediated transport, protein localization and nucleotide binding (Fig. 6). These were also the most highly enriched clusters when looking specifically at proteins identified in ProK treated virions. The cluster cell adhesion was also relatively highly enriched when looking at all proteins, but less so in ProK treated virions as would be expected.


Cellular proteins associated with the interior and exterior of vesicular stomatitis virus virions.

Moerdyk-Schauwecker M, Hwang SI, Grdzelishvili VZ - PLoS ONE (2014)

Functional enrichment analysis.Enrichment analysis and clustering based on gene ontology terms was conducted for (a) all proteins identified and (b) proteins identified in the proteinase K (ProK) treated virions, as described in the materials in methods. The five clusters in each database with the highest enrichment score are depicted here. Numbers above the bars indicate the number of identified proteins associated with each cluster.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4126742&req=5

pone-0104688-g006: Functional enrichment analysis.Enrichment analysis and clustering based on gene ontology terms was conducted for (a) all proteins identified and (b) proteins identified in the proteinase K (ProK) treated virions, as described in the materials in methods. The five clusters in each database with the highest enrichment score are depicted here. Numbers above the bars indicate the number of identified proteins associated with each cluster.
Mentions: To obtain an overview of the types of proteins most commonly associated with purified VSV virions, an enrichment analysis was conducted using gene ontology terms. This analysis was done for all three GO databases (biological process, cellular component and molecular function), first using all the host proteins found in our analysis then looking specifically at proteins associated with ProK treated virions (Fig. 6), as this is the highest purity sample and excludes proteins found exclusively on the exterior of the virion. Related enriched terms were then clustered together to give an overall enrichment score. The most highly enriched clusters, when looking at all the proteins, were vesicles and vesicle mediated transport, protein localization and nucleotide binding (Fig. 6). These were also the most highly enriched clusters when looking specifically at proteins identified in ProK treated virions. The cluster cell adhesion was also relatively highly enriched when looking at all proteins, but less so in ProK treated virions as would be expected.

Bottom Line: Most of these proteins have not been previously shown to be associated with VSV.Functional enrichment analysis indicated the most overrepresented categories were proteins associated with vesicles, vesicle-mediated transport and protein localization.Our study provides a valuable inventory of virion-associated host proteins for further investigation of their roles in the replication cycle, pathogenesis and immunoreactivity of VSV.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Sciences, University of North Carolina at Charlotte, Charlotte, North Carolina, United States of America; Center for Biomedical Engineering and Science, University of North Carolina at Charlotte, Charlotte, North Carolina, United States of America.

ABSTRACT
Virus particles (virions) often contain not only virus-encoded but also host-encoded proteins. Some of these host proteins are enclosed within the virion structure, while others, in the case of enveloped viruses, are embedded in the host-derived membrane. While many of these host protein incorporations are likely accidental, some may play a role in virus infectivity, replication and/or immunoreactivity in the next host. Host protein incorporations may be especially important in therapeutic applications where large numbers of virus particles are administered. Vesicular stomatitis virus (VSV) is the prototypic rhabdovirus and a candidate vaccine, gene therapy and oncolytic vector. Using mass spectrometry, we previously examined cell type dependent host protein content of VSV virions using intact ("whole") virions purified from three cell lines originating from different species. Here we aimed to determine the localization of host proteins within the VSV virions by analyzing: i) whole VSV virions; and ii) whole VSV virions treated with Proteinase K to remove all proteins outside the viral envelope. A total of 257 proteins were identified, with 181 identified in whole virions and 183 identified in Proteinase K treated virions. Most of these proteins have not been previously shown to be associated with VSV. Functional enrichment analysis indicated the most overrepresented categories were proteins associated with vesicles, vesicle-mediated transport and protein localization. Using western blotting, the presence of several host proteins, including some not previously shown in association with VSV (such as Yes1, Prl1 and Ddx3y), was confirmed and their relative quantities in various virion fractions determined. Our study provides a valuable inventory of virion-associated host proteins for further investigation of their roles in the replication cycle, pathogenesis and immunoreactivity of VSV.

Show MeSH
Related in: MedlinePlus