The Src family kinase c-Yes is required for maturation of West Nile virus particles

Alec Hirsch, Guruprasad R. Medigeshi, Heather L. Meyers, Victor De Filippis, Klaus Frueh, Thomas Briese, W. Ian Lipkin, Jay Nelson

Research output: Contribution to journalArticle

61 Citations (Scopus)

Abstract

The role of cellular genes in West Nile virus (WNV) replication is not well understood. Examination of cellular transcripts upregulated during WNV infection revealed an increase in the expression of the src family kinase (SFK) c-Yes. WNV-infected cell lines treated with the SFK inhibitor PP2 demonstrated a 2- to 4-log decrease in viral titers, suggesting that SFK activity is required for completion of the viral replication cycle. RNA interference mediated knock-down of c-Yes, but not c-Src, and similarly reduced virus yield, specifically implicating c-Yes in WNV production. Interestingly, PP2 treatment did not reduce intracellular levels of either viral RNA or protein, suggesting that the drug does not act on the early stages of replication. However, endoglycosidase H (endoH) digestion of the viral envelope (E) glycoprotein revealed that the acquisition of endoH-resistant glycans by E, but not endogenous major histocompatibility complex class I, was reduced in PP2-treated cells, demonstrating that E specifically does not traffic beyond the endoplasmic reticulum in the absence of SFK activity. Electron microscopy further revealed that PP2-treated WNV-infected cells accumulated an increased number of virions in the ER compared to untreated cells. Therefore, we conclude that inhibition of SFK activity did not interfere with virus assembly but prevented transit of virions through the secretory pathway. These results identify c-Yes as a cellular protein that is involved in WNV assembly and egress.

Original languageEnglish (US)
Pages (from-to)11943-11951
Number of pages9
JournalJournal of Virology
Volume79
Issue number18
DOIs
StatePublished - Sep 2005

Fingerprint

West Nile virus
src-Family Kinases
virion
Virion
phosphotransferases (kinases)
Virus Assembly
glycosidases
Glycoside Hydrolases
virus replication
Virus Release
Secretory Pathway
Viral RNA
major histocompatibility complex
Viral Proteins
Virus Diseases
cells
Virus Replication
viral load
RNA Interference
Major Histocompatibility Complex

ASJC Scopus subject areas

  • Immunology

Cite this

The Src family kinase c-Yes is required for maturation of West Nile virus particles. / Hirsch, Alec; Medigeshi, Guruprasad R.; Meyers, Heather L.; De Filippis, Victor; Frueh, Klaus; Briese, Thomas; Lipkin, W. Ian; Nelson, Jay.

In: Journal of Virology, Vol. 79, No. 18, 09.2005, p. 11943-11951.

Research output: Contribution to journalArticle

Hirsch, Alec ; Medigeshi, Guruprasad R. ; Meyers, Heather L. ; De Filippis, Victor ; Frueh, Klaus ; Briese, Thomas ; Lipkin, W. Ian ; Nelson, Jay. / The Src family kinase c-Yes is required for maturation of West Nile virus particles. In: Journal of Virology. 2005 ; Vol. 79, No. 18. pp. 11943-11951.
@article{85063fa196ef4d4aace613d55b43a390,
title = "The Src family kinase c-Yes is required for maturation of West Nile virus particles",
abstract = "The role of cellular genes in West Nile virus (WNV) replication is not well understood. Examination of cellular transcripts upregulated during WNV infection revealed an increase in the expression of the src family kinase (SFK) c-Yes. WNV-infected cell lines treated with the SFK inhibitor PP2 demonstrated a 2- to 4-log decrease in viral titers, suggesting that SFK activity is required for completion of the viral replication cycle. RNA interference mediated knock-down of c-Yes, but not c-Src, and similarly reduced virus yield, specifically implicating c-Yes in WNV production. Interestingly, PP2 treatment did not reduce intracellular levels of either viral RNA or protein, suggesting that the drug does not act on the early stages of replication. However, endoglycosidase H (endoH) digestion of the viral envelope (E) glycoprotein revealed that the acquisition of endoH-resistant glycans by E, but not endogenous major histocompatibility complex class I, was reduced in PP2-treated cells, demonstrating that E specifically does not traffic beyond the endoplasmic reticulum in the absence of SFK activity. Electron microscopy further revealed that PP2-treated WNV-infected cells accumulated an increased number of virions in the ER compared to untreated cells. Therefore, we conclude that inhibition of SFK activity did not interfere with virus assembly but prevented transit of virions through the secretory pathway. These results identify c-Yes as a cellular protein that is involved in WNV assembly and egress.",
author = "Alec Hirsch and Medigeshi, {Guruprasad R.} and Meyers, {Heather L.} and {De Filippis}, Victor and Klaus Frueh and Thomas Briese and Lipkin, {W. Ian} and Jay Nelson",
year = "2005",
month = "9",
doi = "10.1128/JVI.79.18.11943-11951.2005",
language = "English (US)",
volume = "79",
pages = "11943--11951",
journal = "Journal of Virology",
issn = "0022-538X",
publisher = "American Society for Microbiology",
number = "18",

}

TY - JOUR

T1 - The Src family kinase c-Yes is required for maturation of West Nile virus particles

AU - Hirsch, Alec

AU - Medigeshi, Guruprasad R.

AU - Meyers, Heather L.

AU - De Filippis, Victor

AU - Frueh, Klaus

AU - Briese, Thomas

AU - Lipkin, W. Ian

AU - Nelson, Jay

PY - 2005/9

Y1 - 2005/9

N2 - The role of cellular genes in West Nile virus (WNV) replication is not well understood. Examination of cellular transcripts upregulated during WNV infection revealed an increase in the expression of the src family kinase (SFK) c-Yes. WNV-infected cell lines treated with the SFK inhibitor PP2 demonstrated a 2- to 4-log decrease in viral titers, suggesting that SFK activity is required for completion of the viral replication cycle. RNA interference mediated knock-down of c-Yes, but not c-Src, and similarly reduced virus yield, specifically implicating c-Yes in WNV production. Interestingly, PP2 treatment did not reduce intracellular levels of either viral RNA or protein, suggesting that the drug does not act on the early stages of replication. However, endoglycosidase H (endoH) digestion of the viral envelope (E) glycoprotein revealed that the acquisition of endoH-resistant glycans by E, but not endogenous major histocompatibility complex class I, was reduced in PP2-treated cells, demonstrating that E specifically does not traffic beyond the endoplasmic reticulum in the absence of SFK activity. Electron microscopy further revealed that PP2-treated WNV-infected cells accumulated an increased number of virions in the ER compared to untreated cells. Therefore, we conclude that inhibition of SFK activity did not interfere with virus assembly but prevented transit of virions through the secretory pathway. These results identify c-Yes as a cellular protein that is involved in WNV assembly and egress.

AB - The role of cellular genes in West Nile virus (WNV) replication is not well understood. Examination of cellular transcripts upregulated during WNV infection revealed an increase in the expression of the src family kinase (SFK) c-Yes. WNV-infected cell lines treated with the SFK inhibitor PP2 demonstrated a 2- to 4-log decrease in viral titers, suggesting that SFK activity is required for completion of the viral replication cycle. RNA interference mediated knock-down of c-Yes, but not c-Src, and similarly reduced virus yield, specifically implicating c-Yes in WNV production. Interestingly, PP2 treatment did not reduce intracellular levels of either viral RNA or protein, suggesting that the drug does not act on the early stages of replication. However, endoglycosidase H (endoH) digestion of the viral envelope (E) glycoprotein revealed that the acquisition of endoH-resistant glycans by E, but not endogenous major histocompatibility complex class I, was reduced in PP2-treated cells, demonstrating that E specifically does not traffic beyond the endoplasmic reticulum in the absence of SFK activity. Electron microscopy further revealed that PP2-treated WNV-infected cells accumulated an increased number of virions in the ER compared to untreated cells. Therefore, we conclude that inhibition of SFK activity did not interfere with virus assembly but prevented transit of virions through the secretory pathway. These results identify c-Yes as a cellular protein that is involved in WNV assembly and egress.

UR - http://www.scopus.com/inward/record.url?scp=24644497547&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=24644497547&partnerID=8YFLogxK

U2 - 10.1128/JVI.79.18.11943-11951.2005

DO - 10.1128/JVI.79.18.11943-11951.2005

M3 - Article

C2 - 16140770

AN - SCOPUS:24644497547

VL - 79

SP - 11943

EP - 11951

JO - Journal of Virology

JF - Journal of Virology

SN - 0022-538X

IS - 18

ER -