The Ebola Virus Matrix Protein Penetrates into the Plasma Membrane

Adu-Gyamfi, Emmanuel; Soni, Smita P.; Xue, Yi; Digman, Michelle A.; Gratton, Enrico; Stahelin, Robert V.

doi:10.1074/jbc.m112.443960

Cited by 89 publications

(128 citation statements)

References 39 publications

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“…These sequence dissimilarities seem to contribute to the different lipid binding properties of eVP40 and mVP40. Previous studies have demonstrated that eVP40 can associate with vesicles containing PS (20,(22)(23)(24)38). While these previous studies have not discriminated between specific and nonspecific electrostatic interactions with PS, more detailed experiments with PS interactions in cell culture have demonstrated that PS plays an important and specific role in interacting with eVP40 (59,60).…”

Section: Discussionmentioning

confidence: 98%

“…Similarly, proteins that interact with plasma membrane PI(4,5)P 2 may do so with a specific binding motif (35)(36)(37) or through nonspecific electrostatic interactions (30,31). Not surprisingly, several viral matrix proteins, including eVP40 (20,(22)(23)(24)(25)38), have been shown to associate with anionic lipids, such as PS and PI(4,5)P 2 , in vitro and in cells (39)(40)(41)(42)(43)(44)(45)(46), which is important in mediating their assembly and budding.…”

mentioning

confidence: 99%

“…mVP40 and eVP40 are 34% identical (49% homologous) in amino acid sequence, which suggests they may have some different interactions with host cell proteins or lipids. eVP40 harbors an N-terminal domain (NTD) involved in oligomerization (16)(17)(18)(19) and a C-terminal domain (CTD) reported to mediate membrane binding (20)(21)(22)(23)(24)(25). Recent structural analyses of eVP40 (21) and mVP40 (26) have revealed an NTD interface that mediates dimerization, which is required for VP40 plasma membrane localization and budding.…”

mentioning

confidence: 99%

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Investigation of the Lipid Binding Properties of the Marburg Virus Matrix Protein VP40

Wijesinghe

Stahelin

2016

J Virol

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Marburg virus (MARV), which belongs to the virus family Filoviridae, causes hemorrhagic fever in humans and nonhuman primates that is often fatal. MARV is a lipid-enveloped virus that during the replication process extracts its lipid coat from the plasma membrane of the host cell it infects. MARV carries seven genes, one of which encodes its matrix protein VP40 (mVP40), which regulates the assembly and budding of the virions. Currently, little information is available on mVP40 lipid binding properties. Here, we have investigated the in vitro and cellular mechanisms by which mVP40 associates with lipid membranes. mVP40 associates with anionic membranes in a nonspecific manner that is dependent upon the anionic charge density of the membrane. These results are consistent with recent structural determination of mVP40, which elucidated an mVP40 dimer with a flat and extensive cationic lipid binding interface. IMPORTANCEMarburg virus (MARV) is a lipid-enveloped filamentous virus from the family Filoviridae. MARV was discovered in 1967, and yet little is known about how its seven genes are used to assemble and form a new viral particle in the host cell it infects. The MARV matrix protein VP40 (mVP40) underlies the inner leaflet of the virus and regulates budding from the host cell plasma membrane. In vitro and cellular assays in this study investigated the mechanism by which mVP40 associates with lipids. The results demonstrate that mVP40 interactions with lipid vesicles or the inner leaflet of the plasma membrane are electrostatic but nonspecific in nature and are dependent on the anionic charge density of the membrane surface. Small molecules that can disrupt lipid trafficking or reduce the anionic charge of the plasma membrane interface may be useful in inhibiting assembly and budding of MARV. Marburg virus (MARV) and Ebola virus (EBOV) are members of the virus family Filoviridae that are characterized by their filamentous lipid -enveloped morphology (1). Electron microscopy studies revealed that MARV particles have a uniform diameter of approximately 80 nm, with an average particle length of 740 nm (2, 3). MARV harbors a negative-sense RNA genome 19.1 kb in length that includes seven genes (4). The glycoprotein (GP) is a transmembrane protein present in the viral envelope, and it mediates viral entry into the host cell. VP40 is the major matrix protein (mVP40) and completely underlies the lipid envelope of the virus. VP40 alone is sufficient to produce virus-like particles (VLPs) from mammalian cells that are nearly indistinguishable from virions (5-8). In addition to interacting with the host cell membrane and mediating budding, VP40 interacts with the nucleocapsid, which consists of nucleoprotein, VP24, VP30, VP35, and the L protein (9). VP40 also plays other essential roles, such as immunosuppression (10), regulation of viral transcription, and/or replication (11,12), and is also able to interact with GP, coexpression of which leads to GP enrichment at sites of mVP40 budding (13). mVP40 is a peripheral ...

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Section: Discussionmentioning

confidence: 98%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Investigation of the Lipid Binding Properties of the Marburg Virus Matrix Protein VP40

Wijesinghe

Stahelin

2016

J Virol

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“…While VP40 has been shown to associate with lipid vesicles or model membrane systems harboring phosphatidylserine (PS) in vitro (24)(25)(26)(27)(28), little information is available on how VP40 assembles and buds from the plasma membrane of human cells and on what the targets in these processes might be for antiviral intervention.…”

mentioning

confidence: 99%

“…To this end, VP40 has served as an excellent model to investigate Ebola virus budding in vitro. VP40 has been shown to form a high-affinity dimer (21), which is able to transform into hexameric filaments that can concentrate at the inner leaflet of the plasma membrane (21)(22)(23)(24).…”

mentioning

confidence: 99%

Host Cell Plasma Membrane Phosphatidylserine Regulates the Assembly and Budding of Ebola Virus

Adu-Gyamfi

Johnson

Fraser

et al. 2015

J Virol

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199

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Lipid-enveloped viruses replicate and bud from the host cell where they acquire their lipid coat. Ebola virus, which buds from the plasma membrane of the host cell, causes viral hemorrhagic fever and has a high fatality rate. To date, little has been known about how budding and egress of Ebola virus are mediated at the plasma membrane. We have found that the lipid phosphatidylserine (PS) regulates the assembly of Ebola virus matrix protein VP40. VP40 binds PS-containing membranes with nanomolar affinity, and binding of PS regulates VP40 localization and oligomerization on the plasma membrane inner leaflet. Further, alteration of PS levels in mammalian cells inhibits assembly and egress of VP40. Notably, interactions of VP40 with the plasma membrane induced exposure of PS on the outer leaflet of the plasma membrane at sites of egress, whereas PS is typically found only on the inner leaflet. Taking the data together, we present a model accounting for the role of plasma membrane PS in assembly of Ebola virus-like particles. IMPORTANCEThe lipid-enveloped Ebola virus causes severe infection with a high mortality rate and currently lacks FDA-approved therapeutics or vaccines. Ebola virus harbors just seven genes in its genome, and there is a critical requirement for acquisition of its lipid envelope from the plasma membrane of the human cell that it infects during the replication process. There is, however, a dearth of information available on the required contents of this envelope for egress and subsequent attachment and entry. Here we demonstrate that plasma membrane phosphatidylserine is critical for Ebola virus budding from the host cell plasma membrane. This report, to our knowledge, is the first to highlight the role of lipids in human cell membranes in the Ebola virus replication cycle and draws a clear link between selective binding and transport of a lipid across the membrane of the human cell and use of that lipid for subsequent viral entry. Lipid-enveloped viruses harbor a lipid membrane bilayer derived from their host cell during the budding process. This envelope provides the virus stability, protection of its genetic contents, and a reservoir for its transmembrane glycoprotein, which mediates entry into cells (1, 2). The viral lipid envelope may be a viable target for drug development, as particular alterations in the lipid coat or receptor-lipid interaction can inhibit viral entry (3-6). The lipid-dependent budding and egress of some lipid-enveloped viruses have been investigated. For example, it is well established that HIV-1 binds and utilizes 1,2-dioleoyl-sn-glycero-3-phospho-(1=-myo-inositol-4=,5=-bisphosphate) [PI(4,5)P 2 ] enriched in the plasma membrane (PM) inner leaflet for assembly and egress from the cell (7,8). Enteroviruses and flaviviruses use a phosphatidylinositol-4-phosphate [PI(4)P]-enriched organelle to replicate (9), and enteroviruses are packaged into phosphatidylserine (PS)-enriched vesicles, thereby enhancing the efficiency of viral transmission (10). The budding and egres...

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Conformational plasticity of the Ebola virus matrix protein

2014

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Filoviruses are the causative agents of a severe and often fatal hemorrhagic fever with repeated outbreaks in Africa. They are negative sense single stranded enveloped viruses that can cross species barriers from its natural host bats to primates including humans. The small size of the genome poses limits to viral adaption, which may be partially overcome by conformational plasticity. Here we review the different conformational states of the Ebola virus (EBOV) matrix protein VP40 that range from monomers, to dimers, hexamers, and RNA-bound octamers. This conformational plasticity that is required for the viral life cycle poses a unique opportunity for development of VP40 specific drugs. Furthermore, we compare the structure to homologous matrix protein structures from Paramyxoviruses and Bornaviruses and we predict that they do not only share the fold but also the conformational flexibility of EBOV VP40.

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The Ebola Virus Matrix Protein Penetrates into the Plasma Membrane

Cited by 89 publications

References 39 publications

Investigation of the Lipid Binding Properties of the Marburg Virus Matrix Protein VP40

Investigation of the Lipid Binding Properties of the Marburg Virus Matrix Protein VP40

Host Cell Plasma Membrane Phosphatidylserine Regulates the Assembly and Budding of Ebola Virus

Conformational plasticity of the Ebola virus matrix protein

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