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A20/TNFAIP3 inhibits NF-κB activation induced by the Kaposi’s sarcoma-associated herpesvirus vFLIP oncoprotein

Oncogene volume 32, pages 1223–1232 (2013)Cite this article

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Abstract

Kaposi’s sarcoma-associated herpesvirus (KSHV) K13/vFLIP (viral Flice-inhibitory protein) induces transcription of numerous genes through NF-κB activation, including pro-inflammatory cytokines, which contribute to the pathogenesis of Kaposi’s sarcoma (KS). In this study, we report that KSHV vFLIP induces the expression of the NF-κB regulatory proteins A20, ABIN-1 and ABIN-3 (A20-binding NF-κB inhibitors) in primary human endothelial cells, and that KS spindle cells express A20 in KS tissue. In reporter assays, A20 strongly impaired vFLIP-induced NF-κB activation in 293T cells, but ABIN-1 and ABIN-3 did not. Mutational analysis established that the C-terminal domain (residues 427–790) is critical for A20 modulation of NF-κB, but the ubiquitin-editing OTU (ovarian tumor) domain is not. In functional assays, A20 inhibited vFLIP-induced expression of the chemokine IP-10, reduced vFLIP-induced cell proliferation and increased IKK1 protein levels. Thus, we demonstrate that A20 negatively regulates NF-κB activation directly induced by KSHV vFLIP. By attenuating excessive and prolonged vFLIP-induced NF-κB activation that could be harmful to KSHV-infected cells, A20 likely has an important role in the pathogenesis of KSHV-associated diseases, in which vFLIP is expressed.

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Acknowledgements

We thank G Nolan (Stanford University), BK Weaver (Missouri State University), J-M Peloponese Jr, K-T Jeang (NIAID, NIH), DB Conze, JD Ashwell (NCI, NIH), AV Grishin (University of Southern California), A Leonardi (University of Naples, Italy), R Lin (Jewish General Hospital, Canada) for reagents; the patients who volunteered for this study; Dr MN Polizzotto (NCI, NIH), K Aleman and K Wyvill for patient care; the Confocal Microscopy Core Facility (Laboratory of Experimental Carcinogenesis, CCR, NCI) for providing LSM510 and the DNA Minicore Facility (Laboratory of Experimental Carcinogenesis, CCR, NCI) for DNA sequencing, and K Sakakibara (NCI, NIH) for her help in reporter assay. We also thank Drs D Lowy, O Salvucci, H Ohnuki and all members of Laboratory of Cellular Oncology Branch for their advice and help on this work. This study was supported by the intramural research program in Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD20892, USA.

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  1. S Sakakibara and G Espigol-Frigole: These authors contributed equally to this work

Authors and Affiliations

  1. Laboratory of Cellular Oncology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA

    S Sakakibara, G Espigol-Frigole, P Gasperini & G Tosato

  2. HIV and AIDS Malignancy Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA

    T S Uldrick & R Yarchoan

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  1. S Sakakibara
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Sakakibara, S., Espigol-Frigole, G., Gasperini, P. et al. A20/TNFAIP3 inhibits NF-κB activation induced by the Kaposi’s sarcoma-associated herpesvirus vFLIP oncoprotein. Oncogene 32, 1223–1232 (2013). https://doi.org/10.1038/onc.2012.145

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