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  • Original Paper
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A novel dominant-negative mutant form of Epstein–Barr virus latent membrane protein-1 (LMP1) selectively and differentially impairs LMP1 and TNF signaling pathways

Oncogene volume 23, pages 2681–2693 (2004)Cite this article

A Corrigendum to this article was published on 21 October 2004

Abstract

The latent membrane protein-1 (LMP1) is an integral membrane molecule expressed by Epstein–Barr virus (EBV) during viral latency and displays properties of a constitutively activated member of the TNF receptor family. LMP1 is required for B-cell or monocyte immortalization induced by EBV and is sufficient to transform rodent fibroblasts. Transforming potential of LMP1 is mediated by its cytoplasmic C-terminal domain, which activates various cellular signaling pathways including NFκB and JNK. In this report, we constructed mutants of LMP1 with preserved membrane spanning domain but mutated in the C-terminal domain and a second truncated C-terminal LMP1 fused to the enhanced green fluorescent protein. This latter mutant, termed LMP1-CT, impairs signaling by ectopic LMP1 as well as endogenous EBV-expressed wild-type (wt) LMP1. In contrast to dominant-negative mutants of LMP1 with preserved membrane spanning domains, LMP1-CT was unable to bind wt LMP1 to form an inactive complex. Its dominant-negative effects were due to binding and sequestration of LMP1 adapters TRAF2 and TRADD as assessed by coimmunoprecipitation experiments and confocal analysis. The effect was selective since LMP1-CT did not inhibit IL-1β-induced signaling, whereas it impaired TNF-triggered NFκB and JNK signals without affecting TNF-induced apoptosis. In addition and in contrast to LMP1 constructs with membrane localization, LMP-CT did not display cytostatic properties in noninfected cells. Importantly, LMP1-CT inhibited survival induced by LMP1 in an EBV-transformed T-cell line expressing the type II viral latency commonly found in the majority of EBV-associated human tumors. These data demonstrate that LMP1-CT is a new tool to explore the differences between LMP1 and TNF signaling and may facilitate the design of molecules with potential therapeutic roles.

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Acknowledgements

We thank Jean Luc Baert, Patrick Mehlen, Tularik corporation, Pierre Busson, B Derijard, and M Ptashne for reagents, and Alexis Verger for critical reading of the manuscript. This work was supported by the Association pour la Recherche sur le Cancer (ARC; Grants 9615 and 5455), CNRS, Lille II, Institut Pasteur de Lille, and INSERM.

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Authors and Affiliations

  1. CNRS UMR 8527, Institut de Biologie de Lille, BP 447, Lille Cedex, 59021, France

    Eric Adriaenssens, Alexandra Mougel, Gautier Goormachtigh, Claude Auriault & Jean Coll

  2. Sedac Therapeutics, 59120 Loos, France

    Estelle Loing

  3. CNRS UMR 8117, Institut de Biologie de Lille, BP 447, Lille Cedex, 59021, France

    Véronique Fafeur

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  1. Eric Adriaenssens
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Correspondence to Jean Coll.

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Adriaenssens, E., Mougel, A., Goormachtigh, G. et al. A novel dominant-negative mutant form of Epstein–Barr virus latent membrane protein-1 (LMP1) selectively and differentially impairs LMP1 and TNF signaling pathways. Oncogene 23, 2681–2693 (2004). https://doi.org/10.1038/sj.onc.1207432

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