Lysosome-Associated Membrane Proteins Support the Furin-Mediated Processing of the Mumps Virus Fusion Protein

• Published in: Journal of virology Vol. 94; no. 12
• Main Authors: Ueo, Ayako, Kubota, Marie, Shirogane, Yuta, Ohno, Shinji, Hashiguchi, Takao, Yanagi, Yusuke
• Format: Journal Article
• Language: English
• Published: United States American Society for Microbiology 01.06.2020
• Subjects: A549 Cells; Animals; Cell Membrane - metabolism; Cell Membrane - virology; Chlorocebus aethiops; Furin - genetics; Furin - metabolism; Gene Expression Regulation; Giant Cells - chemistry; Giant Cells - metabolism; HEK293 Cells; HeLa Cells; HN Protein - genetics; HN Protein - metabolism; Host-Pathogen Interactions - genetics; Humans; Lysosomal Membrane Proteins - genetics; Lysosomal Membrane Proteins - metabolism; Lysosomal-Associated Membrane Protein 2 - genetics; Lysosomal-Associated Membrane Protein 2 - metabolism; Lysosomes - metabolism; Lysosomes - virology; Mumps virus - genetics; Mumps virus - metabolism; Neoplasm Proteins - genetics; Neoplasm Proteins - metabolism; Protein Binding; Proteolysis; Signal Transduction; Spotlight; Vero Cells; Viral Fusion Proteins - genetics; Viral Fusion Proteins - metabolism; Virus Internalization; Virus-Cell Interactions; fusion protein; mumps virus; LAMP; furin
• ISSN: 0022-538X; 1098-5514
• DOI: 10.1128/JVI.00050-20

Mumps virus (MuV), an enveloped RNA virus of the family and the causative agent of mumps, affects the salivary glands and other glandular tissues as well as the central nervous system. The virus enters the cell by inducing the fusion of its envelope with the plasma membrane of the target cell. Membrane fusion is mediated by MuV envelope proteins: the hemagglutinin-neuraminidase and fusion (F) protein. Cleavage of the MuV F protein (MuV-F) into two subunits by the cellular protease furin is a prerequisite for fusion and virus infectivity. Here, we show that 293T (a derivative of HEK293) cells do not produce syncytia upon expression of MuV envelope proteins or MuV infection. This failure is caused by the inefficient MuV-F cleavage despite the presence of functional furin in 293T cells. An expression cloning strategy revealed that overexpression of lysosome-associated membrane proteins (LAMPs) confers on 293T cells the ability to produce syncytia upon expression of MuV envelope proteins. The LAMP family comprises the ubiquitously expressed LAMP1 and LAMP2, the interferon-stimulated gene product LAMP3, and the cell type-specific proteins. The expression level of the LAMP3 gene, but not of LAMP1 and LAMP2 genes, differed markedly between 293T and HEK293 cells. Overexpression of LAMP1, LAMP2, or LAMP3 allowed 293T cells to process MuV-F efficiently. Furthermore, these LAMPs were found to interact with both MuV-F and furin. Our results indicate that LAMPs support the furin-mediated cleavage of MuV-F and that, among them, LAMP3 may be critical for the process, at least in certain cells. The cellular protease furin mediates proteolytic cleavage of many host and pathogen proteins and plays an important role in viral envelope glycoprotein maturation. MuV, an enveloped RNA virus of the family and an important human pathogen, enters the cell through the fusion of its envelope with the plasma membrane of the target cell. Membrane fusion is mediated by the viral attachment protein and the F protein. Cleavage of MuV-F into two subunits by furin is a prerequisite for fusion and virus infectivity. Here, we show that LAMPs support the furin-mediated cleavage of MuV-F. Expression levels of LAMPs affect the processing of MuV-F and MuV-mediated membrane fusion. Among LAMPs, the interferon-stimulated gene product LAMP3 is most critical in certain cells. Our study provides potential targets for anti-MuV therapeutics.