![]() ![]() ( B) 293T cells were co-transfected with HA-tagged ITCH and/or FLAG-tagged Z protein of MOPV (ZMop-FLAG), LASV (ZLas-FLAG), LCMV (ZLcm-FLAG), LUJV (ZLuj-FLAG), JUNV (ZJun-FLAG). ![]() FLAG-tagged Z co-immunoprecipitated proteins were detected by western blotting ( n = 3 independent experiments). After 15 h, cells were lysed and protein extracts were readily analyzed (input) or incubated with anti-HA magnetic beads for ITCH immunoprecipitation (IP (HA)). ( A) 293T cells were co-transfected with HA-tagged ITCH and/or FLAG-tagged Z protein of MOPV (ZMop-FLAG), LASV (ZLas-FLAG), LCMV (ZLcm-FLAG), LUJV (ZLuj-FLAG), JUNV (ZJun-FLAG). ITCH LASV Lassa fever MOPV Z matrix protein egress infection yeast two-hybrid screening.Ĭonfirmation of the interaction between various arenavirus Z proteins and the cellular protein ITCH. Thus, ITCH constitutes a novel interactor of LASV and MOPV Z proteins that is involved in virus assembly and release. More precisely, ITCH was involved in the egress of virus-like particles and the release of infectious progeny viruses. The silencing of ITCH was shown to affect the replication of the old-world mammarenaviruses LASV, MOPV, Lymphocytic choriomeningitis virus (LCMV) and to a lesser extent Lujo virus (LUJV). The PPxY late-domain motif of the Z proteins is required for the interaction with ITCH, although the E3 ubiquitin-ligase activity of ITCH is not involved in Z ubiquitination. The cellular proteins ITCH and WWP1, two members of the Nedd4 family of HECT E3 ubiquitin ligases, were found to bind the Z proteins of LASV, MOPV and other arenaviruses. To better characterize the interactions involved, a yeast two-hybrid screen was conducted using the Z proteins from LASV and MOPV as a bait. The Z matrix protein of arenaviruses is essential to virus assembly and budding by recruiting host factors, a mechanism that remains partially defined. LASV is the causative agent of Lassa fever, a deadly hemorrhagic fever endemic in West Africa, whereas MOPV is non-pathogenic in humans. Lassa virus (LASV) and Mopeia virus (MOPV) are two closely related, rodent-born mammarenaviruses. ![]()
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