NEDD4 family ubiquitin ligases associate with LCMV Z's PPXY domain and are required for virus budding, but not via direct ubiquitination of Z
| Autor: | Bryan A. Ballif, Loan Dang, David J. Shirley, Jamie A. Kelly, Jason Botten, Emily A. Bruce, Ethan B. Mattice, Inessa Manuelyan, Philip Eisenhauer, Marion E. Weir, Joseph P. Klaus, Christopher M. Ziegler, Benjamin R. King |
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| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
Nedd4 Ubiquitin Protein Ligases
viruses Interaction Networks Cell Membranes Virus Replication Biochemistry Virions Ligases Ubiquitin TSG101 Lymphocytic choriomeningitis virus Small interfering RNAs Post-Translational Modification Amino Acids Biology (General) 0303 health sciences biology Chemistry Organic Compounds 030302 biochemistry & molecular biology Intracellular Signaling Peptides and Proteins Virus Release 3. Good health Cell biology Enzymes Nucleic acids Physical Sciences Basic Amino Acids Cellular Structures and Organelles Research Article QH301-705.5 Viral budding Protein domain Immunology macromolecular substances Lymphocytic Choriomeningitis Viral Structure Microbiology ESCRT 03 medical and health sciences Protein Domains Virology Genetics Humans Protein Interaction Domains and Motifs Non-coding RNA Molecular Biology 030304 developmental biology Viral matrix protein Virus Assembly Lysine Organic Chemistry Ubiquitination Chemical Compounds Biology and Life Sciences Proteins Membrane Proteins Cell Biology RC581-607 Ubiquitin Ligases Gene regulation Membrane protein biology.protein Enzymology RNA Parasitology Gene expression Immunologic diseases. Allergy |
| Zdroj: | PLoS Pathogens, Vol 15, Iss 11, p e1008100 (2019) PLOS Pathogens PLoS Pathogens |
| ISSN: | 1553-7374 1553-7366 |
| Popis: | Viral late domains are used by many viruses to recruit the cellular endosomal sorting complex required for transport (ESCRT) to mediate membrane scission during viral budding. Unlike the P(S/T)AP and YPX(1–3)L late domains, which interact directly with the ESCRT proteins Tsg101 and ALIX, the molecular linkage connecting the PPXY late domain to ESCRT proteins is unclear. The mammarenavirus lymphocytic choriomeningitis virus (LCMV) matrix protein, Z, contains only one late domain, PPXY. We previously found that this domain in LCMV Z, as well as the ESCRT pathway, are required for the release of defective interfering (DI) particles but not infectious virus. To better understand the molecular mechanism of ESCRT recruitment by the PPXY late domain, affinity purification-mass spectrometry was used to identify host proteins that interact with the Z proteins of the Old World mammarenaviruses LCMV and Lassa virus. Several Nedd4 family E3 ubiquitin ligases interact with these matrix proteins and in the case of LCMV Z, the interaction was PPXY-dependent. We demonstrated that these ligases directly ubiquitinate LCMV Z and mapped the specific lysine residues modified. A recombinant LCMV containing a Z that cannot be ubiquitinated maintained its ability to produce both infectious virus and DI particles, suggesting that direct ubiquitination of LCMV Z alone is insufficient for recruiting ESCRT proteins to mediate virus release. However, Nedd4 ligases appear to be important for DI particle release suggesting that ubiquitination of targets other than the Z protein itself is required for efficient viral ESCRT recruitment. Author summary Enveloped viruses derive their lipid bilayer from either the cellular plasma membrane or an intracellular organelle during the process of viral budding in which a virus particle is formed at a membrane. Many enveloped viruses recruit the cellular endosomal sorting complex required for transport (ESCRT) in order to efficiently cut the membrane that connects a newly budded, but not released, virus particle from its parent membrane. Late domains, which are short protein motifs found in numerous enveloped viruses, specifically recruit ESCRT for this process. Two types of late domains accomplish this by binding directly to ESCRT proteins. A third late domain, PPXY, recruits ESCRT proteins through an unknown, indirect linkage. In this study, we sought to identify proteins that may bridge the PPXY late domain and ESCRT proteins. We found that Nedd4 family ubiquitin ligases interact with the PPXY domain in the mammarenavirus Z protein resulting in ubiquitination of Z at two lysine residues. However, Z ubiquitination was largely dispensable for the virus. Conversely, Nedd4 ubiquitin ligases were critical during infection suggesting that the most important contribution made to virus release by Nedd4 ligases is not direct ubiquitination of the viral matrix protein, but possibly the ubiquitination of cellular proteins or other viral proteins. |
| Databáze: | OpenAIRE |
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