Lysosome-Associated Membrane Proteins Support the Furin-Mediated Processing of the Mumps Virus Fusion Protein
Lysosome-Associated Membrane Proteins Support the Furin-Mediated Processing of the Mumps Virus Fusion Protein
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DOI：
10.1128/jvi.00050-20
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发表时间：
2020-04
影响因子：
5.4
通讯作者：
Ayako Ueo;M. Kubota;Yuta Shirogane;S. Ohno;T. Hashiguchi;Y. Yanagi
中科院分区：
文献类型：
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作者：
Ayako Ueo;M. Kubota;Yuta Shirogane;S. Ohno;T. Hashiguchi;Y. Yanagi
细胞蛋白酶弗林蛋白酶介导许多宿主和病原体蛋白的蛋白水解切割，并在病毒包膜糖蛋白成熟中起重要作用。MuV是副粘病毒科的一种有包膜的RNA病毒，是人类重要的病原体，通过其包膜与靶细胞的质膜融合进入细胞。膜融合由病毒附着蛋白和F蛋白介导。通过弗林蛋白酶将MuV-F切割成两个亚基是融合和病毒感染性的先决条件。在这里，我们表明，LAMP支持弗林蛋白酶介导的切割MuV-F。LAMPs的表达水平影响MuV-F和MuV介导的膜融合的过程。在LAMP中，干扰素刺激的基因产物LAMP 3在某些细胞中是最关键的。我们的研究为抗MuV治疗提供了潜在的靶点。摘要腮腺炎病毒（MuV）是副粘病毒科的一种有包膜的RNA病毒，是腮腺炎的病原体，影响唾液腺和其他腺组织以及中枢神经系统。病毒通过诱导其包膜与靶细胞的质膜融合而进入细胞。膜融合由MuV包膜蛋白介导：血凝素-神经氨酸酶和融合（F）蛋白。MuV F蛋白（MuV-F）被细胞蛋白酶弗林蛋白酶切割成两个亚基是融合和病毒感染性的先决条件。在这里，我们表明，293 T（HEK 293的衍生物）细胞不产生合胞体表达的MuV包膜蛋白或MuV感染。这种失败是由于MuV-F切割效率低下引起的，尽管293 T细胞中存在功能性弗林蛋白酶。表达克隆策略揭示，溶酶体相关膜蛋白（LAMP）的过表达赋予293 T细胞在表达MuV包膜蛋白后产生合胞体的能力。LAMP家族包括广泛表达的LAMP 1和LAMP 2、干扰素刺激的基因产物LAMP 3和细胞类型特异性蛋白。在293 T和HEK 293细胞中，LAMP 3基因的表达水平显著不同，而LAMP 1和LAMP 2基因的表达水平无显著差异。LAMP 1、LAMP 2或LAMP 3的过表达允许293 T细胞有效地加工MuV-F。此外，这些LAMP被发现与MuV-F和弗林蛋白酶两者相互作用。我们的研究结果表明，LAMPs支持弗林蛋白酶介导的MuV-F裂解，其中，LAMP 3可能是关键的过程中，至少在某些细胞。细胞蛋白酶弗林蛋白酶介导许多宿主和病原体蛋白的蛋白水解裂解，并在病毒包膜糖蛋白成熟中起重要作用。MuV是副粘病毒科的一种有包膜的RNA病毒，是人类重要的病原体，通过其包膜与靶细胞的质膜融合进入细胞。膜融合由病毒附着蛋白和F蛋白介导。通过弗林蛋白酶将MuV-F切割成两个亚基是融合和病毒感染性的先决条件。在这里，我们表明，LAMP支持弗林蛋白酶介导的切割MuV-F。LAMPs的表达水平影响MuV-F和MuV介导的膜融合的过程。在LAMP中，干扰素刺激的基因产物LAMP 3在某些细胞中是最关键的。我们的研究为抗MuV治疗提供了潜在的靶点。
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 Paramyxoviridae 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. ABSTRACT Mumps virus (MuV), an enveloped RNA virus of the Paramyxoviridae 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. IMPORTANCE 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 Paramyxoviridae 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.