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Adenovirus 5 E1A is responsible for increased expression of insulin receptor substrate 4 in established adenovirus 5-transformed cell lines and interacts with IRS components activating the PI3 kinase/Akt signalling pathway

Oncogene volume 28, pages 686–697 (2009)Cite this article

Abstract

Using mass spectrometric analysis insulin receptor substrate 4 (IRS-4) has been identified as a novel adenovirus 5 early region 1A (Ad5E1A)-binding protein. IRS-4 interacts with both the transcriptional activation domain (conserved region 3) and the N-terminal region of Ad5E1A13S. Prolonged expression of Ad5E1A13S is required for the observed dramatic increase in the levels of IRS-4 mRNA and protein in Ad5E1-transformed human cell lines. Once expressed, as well as binding to E1A and the insulin receptor, IRS-4 remains tyrosine phosphorylated and constitutively associates with the regulatory p85 subunit of phosphoinositide 3 kinase, resulting in the phosphorylation of Akt (causing activation) and GSK-3β (causing inhibition). Reducing IRS-4 expression using small interfering RNA (siRNA) in established Ad5E1A-expressing cell lines decreases the activation of Akt and cellular proliferation. During Ad5 infection, IRS-4 is not expressed. However, Ad5E1A associates with IRS-1, increasing Akt and GSK-3β phosphorylation and tyrosine phosphorylation of IRS-1 itself. We conclude that the association and altered regulation of IRS proteins by Ad5E1A contribute to the adenovirus-transformed phenotype and modulates viral infection in an Akt-dependent manner.

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Acknowledgements

We are most grateful to Dr Joe Mymryk (University of Western Ontario) for the gift of various AdE1A constructs, Professor Thomas Dobner (University of Hamburg) for a gift of the Ad5-transformed rat cell lines, Professor Derek Leroith (Mount Sinai School of Medicine) for IRS-4 cDNA and Cancer Research UK for their financial support.

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Authors and Affiliations

  1. Cancer Research UK School of Cancer Sciences, University of Birmingham, Birmingham, West Midlands, UK

    N J Shimwell, A Martin, R K Bruton, A N Blackford, G G Sedgwick, P H Gallimore, A S Turnell & R J A Grand

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  1. N J Shimwell
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  2. A Martin
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  3. R K Bruton
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Correspondence to R J A Grand.

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Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc)

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Shimwell, N., Martin, A., Bruton, R. et al. Adenovirus 5 E1A is responsible for increased expression of insulin receptor substrate 4 in established adenovirus 5-transformed cell lines and interacts with IRS components activating the PI3 kinase/Akt signalling pathway. Oncogene 28, 686–697 (2009). https://doi.org/10.1038/onc.2008.417

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