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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. 2004 Apr 08; 23(15):2681-93.O

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 NFkappaB 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-1beta-induced signaling, whereas it impaired TNF-triggered NFkappaB 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.

Authors+Show Affiliations

CNRS UMR 8527, Institut de Biologie de Lille, BP 447, 59021 Lille Cedex, France.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

14767477

Citation

Adriaenssens, Eric, 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, vol. 23, no. 15, 2004, pp. 2681-93.
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. 2004;23(15):2681-93.
Adriaenssens, E., Mougel, A., Goormachtigh, G., Loing, E., Fafeur, V., Auriault, C., & Coll, J. (2004). 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(15), 2681-93.
Adriaenssens E, 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. 2004 Apr 8;23(15):2681-93. PubMed PMID: 14767477.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - A novel dominant-negative mutant form of Epstein-Barr virus latent membrane protein-1 (LMP1) selectively and differentially impairs LMP1 and TNF signaling pathways. AU - Adriaenssens,Eric, AU - Mougel,Alexandra, AU - Goormachtigh,Gautier, AU - Loing,Estelle, AU - Fafeur,Véronique, AU - Auriault,Claude, AU - Coll,Jean, PY - 2004/2/10/pubmed PY - 2004/5/27/medline PY - 2004/2/10/entrez SP - 2681 EP - 93 JF - Oncogene JO - Oncogene VL - 23 IS - 15 N2 - 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 NFkappaB 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-1beta-induced signaling, whereas it impaired TNF-triggered NFkappaB 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. SN - 0950-9232 UR - https://www.unboundmedicine.com/medline/citation/14767477/A_novel_dominant_negative_mutant_form_of_Epstein_Barr_virus_latent_membrane_protein_1__LMP1__selectively_and_differentially_impairs_LMP1_and_TNF_signaling_pathways_ L2 - https://doi.org/10.1038/sj.onc.1207432 DB - PRIME DP - Unbound Medicine ER -