Mapping the interactome of HPV E6 and E7 oncoproteins with the Ubiquitin-Proteasome System.

Fiche publication

Date publication

août 2017

Journal

The FEBS journal

Auteurs

Membres identifiés du Cancéropôle Est :
Dr MASSON Murielle , Dr TRAVE Gilles , Dr ZANIER Katia , Dr NOMINE Yves

Tous les auteurs :
Poirson J, Biquand E, Straub ML, Cassonnet P, Nominé Y, Jones L, van der Werf S, Travé G, Zanier K, Jacob Y, Demeret C, Masson M

Lien Pubmed

https://www.ncbi.nlm.nih.gov/pubmed/28786561

Résumé

Protein ubiquitination and its reverse reaction, deubiquitination, regulate protein stability, protein binding activity and their subcellular localization. These reactions are catalyzed by the enzymes E1, E2 and E3 Ubiquitin ligases and deubiquitinases. The ubiquitin-proteasome system is targeted by viruses for the sake of their replication and to escape host immune response. To identify novel partners of HPV16 E6 and E7 proteins, we assembled and screened a library of 590 cDNAs related to the UPS by using the Gaussia princeps luciferase Protein Complementation Assay. HPV16 E6 was found to bind to the HECT-type Ubiquitin ligase (E6AP), three RING-type Ubiquitin ligases (MGRN1, LNX3, LNX4) and the deubiquitinase USP15. Except for E6AP, the binding of UPS factors did not require the LxxLL-binding pocket of HPV16 E6. LNX3 bound preferentially to all high-risk mucosal HPV E6 tested whereas LNX4 bound specifically to HPV16 E6. HPV16 E7 was found to bind to several BTB domain-containing proteins (such as TNFAIP1/KCTD13) that are potential substrate adaptors of Cullin 3-RING Ubiquitin ligases, to RING-type Ubiquitin ligases implicated in innate immunity (RNF135, TRIM32, TRAF2, TRAF5), to the substrate adaptor DCAF15 of Cullin 4-RING Ubiquitin ligase and to some deubiquitinases (USP29, USP33). The binding to UPS factors did not require the LxCxE motif but rather the C-terminus region of HPV16 E7 protein. The identified UPS factors interacted with most of E7 proteins across different HPV types. This study establishes a strategy for the rapid identification of interactions between host or pathogen proteins and the human ubiquitination system. This article is protected by copyright. All rights reserved.