A novel method for screening viral interferon-resistance genes

Clarke, Daniel T. W., Irving, Aaron T., Lambley, Eleanore H., Payne, Elizabeth and McMillan, Nigel A.J. (2004) A novel method for screening viral interferon-resistance genes. Journal of Interferon And Cytokine Research, 24 8: 470-477.


Author(s) Clarke, Daniel T. W.
Irving, Aaron T.
Lambley, Eleanore H.
Payne, Elizabeth
McMillan, Nigel A.J.
Title A novel method for screening viral interferon-resistance genes
Journal name Journal of Interferon And Cytokine Research
Publication date 2004
Volume number 24
Issue number 8
ISSN 1079-9907
Start page 470
End page 477
Total pages 8
Place of publication Larchmont
Publisher Mary Ann Liebert Inc Publ
Collection year 2004
Language eng
Subject 320000 Medical and Health Sciences
C1
320402 Medical Virology
730108 Cancer and related disorders
Abstract Many viruses have evolved mechanisms to antagonize the interferon (IFN) system, targeting all the major components involved in receptor binding and signaling. Although a number of these vital proteins are homologous to cellular proteins involved in IFN downregulation (e.g., viral IFN regulatory factors [vIRFs]), many share little resemblance to known proteins. To determine the IFN-blocking properties of these proteins, functional assays are required. Here, we present a new and rapid functional screening method, based on the 2fTGH cell line, which is able to determine viral gene products that inhibit the IFN-alpha/Jak-Stat signaling pathway. Expression cloning of viral IFN-blocking genes into 2fTGH and consequent selection with IFN-alpha and 6-thioguanine result in the outgrowth of cells that are no longer responsive to IFN-alpha. We also demonstrate that selection occurs if members of the Jak-Stat signaling pathway are lost. To show the utility of our system, we have used a known suppressor of IFN signaling, the human papillomavirus (HPV) E7 gene. Expression of E7 causes the loss of ability of 2fTGH cells to respond to IFN-alpha treatment because of a functional disruption of the signaling pathway. This approach offers a new strategy for identifying novel viral genes or new functions of already described viral genes that have a role in IFN-alpha signaling inhibition.
Keyword(s) Biochemistry & Molecular Biology
Cell Biology
Immunology
Transcription Factor Isgf-3
Nonstructural Protein Nss
Signal-transduction
Stimulated Transcription
Regulatory Factor
Alpha Interferon
I Interferons
Cell-line
Receptor
Activation
 
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http://dx.doi.org/10.1089/1079990041689610  
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