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  • Original Article
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Genetic deletion of PKR abrogates TNF-induced activation of IκBα kinase, JNK, Akt and cell proliferation but potentiates p44/p42 MAPK and p38 MAPK activation

Oncogene volume 26, pages 1201–1212 (2007)Cite this article

Abstract

Double-stranded RNA-dependent protein kinase (PKR), a ubiquitously expressed serine/threonine kinase, has been implicated in the regulation or modulation of cell growth through multiple signaling pathways, but how PKR regulates tumor necrosis factor (TNF)-induced signaling pathways is poorly understood. In the present study, we used fibroblasts derived from PKR gene-deleted mice to investigate the role of PKR in TNF-induced activation of nuclear factor-κB (NF-κB), mitogen-activated protein kinases (MAPKs) and growth modulation. We found that in wild-type mouse embryonic fibroblast (MEF), TNF induced NF-κB activation as measured by DNA binding but deletion of PKR abolished this activation. This inhibition was associated with suppression of inhibitory subunit of NF-κB (IκB)α kinase (IKK) activation, IκBα phosphorylation and degradation, p65 phosphorylation and nuclear translocation, and NF-κB-dependent reporter gene transcription. TNF-induced Akt activation needed for IKK activation was also abolished by deletion of PKR. NF-κB activation was diminished in PKR-deleted cells transfected with TNF receptor (TNFR) 1, TNFR-associated death domain and TRAF2 plasmids; NF-κB activated by NF-κB-inducing kinase, IKK or p65, however, was minimally affected. Among the MAPKs, it was interesting that whereas TNF-induced c-Jun N-terminal kinase (JNK) activation was abolished, activation of p44/p42 MAPK and p38 MAPK was potentiated in PKR-deleted cells. TNF induced the expression of NF-κB-regulated gene products cyclin D1, c-Myc, matrix metalloproteinase-9, survivin, X-linked inhibitor-of-apoptosis protein (IAP), IAP1, Bcl-xL, A1/Bfl-1 and Fas-associated death domain protein-like IL-1β-converting enzyme-inhibitory protein in wild-type MEF but not in PKR−/− cells. Similarly, TNF induced the proliferation of wild-type cells, but this proliferation was completely suppressed in PKR-deleted cells. Overall, our results indicate that PKR differentially regulates TNF signaling; IKK, Akt and JNK were positively regulated, whereas p44/p42 MAPK and p38 MAPK were negatively regulated.

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Abbreviations

dsRNA:

double-stranded RNA

PKR:

dsRNA-dependent protein kinase

NF-κB:

nuclear factor-κB

IκB:

inhibitory subunit of NF-κB

IKK:

IκBα kinase

SEAP:

secretory alkaline phosphatase

IAP:

inhibitor-of-apoptosis protein

IL:

interleukin

FADD:

Fas-associated death domain protein

FLICE:

FADD-like IL-1β-converting enzyme

FLIP:

FLICE-inhibitory protein

MMP:

matrix metalloproteinase

LPS:

lipopolysaccharide

JNK:

c-Jun N-terminal kinase

MAPK:

mitogen-activated protein kinase

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Acknowledgements

We thank Ann Sutton for carefully proofreading the manuscript and providing valuable comments. Dr Takada is an Odyssey Program Special Fellow at The University of Texas MD Anderson Cancer Center. Dr Aggarwal is a Ransom Horne Jr, Professor of Cancer Research.

This work was supported by a grant from the Clayton Foundation for Research (to BBA), Department of Defense US Army Breast Cancer Research Program Grant BC010610 (to BBA), National Institutes of Health PO1 Grant CA91844 on lung chemoprevention (to BBA), National Institutes of Health P50 Head and Neck SPORE Grant P50CA97007 (to BBA) and grants from the Odyssey Program and the Theodore N Law Award for Scientific Achievement Fund from The University of Texas MD Anderson Cancer Center (to YT).

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  1. Department of Experimental Therapeutics, Cytokine Research Laboratory, The University of Texas MD Anderson Cancer Center, Houston, TX, USA

    Y Takada, H Ichikawa & B B Aggarwal

  2. Department of Thoracic and Cardiovascular Surgery, The University of Texas MD Anderson Cancer Center, Houston, TX, USA

    A Pataer & S Swisher

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Takada, Y., Ichikawa, H., Pataer, A. et al. Genetic deletion of PKR abrogates TNF-induced activation of IκBα kinase, JNK, Akt and cell proliferation but potentiates p44/p42 MAPK and p38 MAPK activation. Oncogene 26, 1201–1212 (2007). https://doi.org/10.1038/sj.onc.1209906

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