The Ebola Virus matrix protein, VP40, requires phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2) for extensive oligomerization at the plasma membrane and viral egress

Johnson, Kristen A.; Taghon, Geoffrey J.; Scott, Jordan L.; Stahelin, Robert V.

doi:10.1038/srep19125

Cited by 79 publications

(207 citation statements)

References 62 publications

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“…Moreover, MARV VP40 acts as an anionic charge sensor (35,40), while EBOV VP40 harbors PS selectivity as shown here and in previous studies (14,27), which further supports PS selective binding properties of the EBOV VP40 CTD.…”

Section: Discussionsupporting

confidence: 86%

“…Alternatively, VP40 is known to harbor important interactions with phosphoinositides at the plasma membrane, which have been shown to stabilize VP40 oligomers (14). Notably, VP40 oligomers are abundant at the plasma membrane interface (12,13) perhaps efficient assembly of VP40 is necessary for phosphoinositide interactions (14) while assembly of L117R is not possible ! !…”

Section: Discussionmentioning

confidence: 99%

“…VP40 is a peripheral membrane protein that regulates budding as well as virus structure and stability (6-8). VP40 is the most abundant protein contained in mature virions (9), and can associate with (10) and oligomerize on (8,(11)(12)(13)(14) the plasma membrane (PM) inner leaflet, where it acquires the viral lipid envelope.…”

Section: Introductionmentioning

confidence: 99%

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A cationic, C-terminal patch and structural rearrangements in Ebola virus matrix VP40 protein control its interactions with phosphatidylserine

Vecchio

Frick

Jeevan

et al. 2018

Journal of Biological Chemistry

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134

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Ebola virus (EBOV) is a filamentous lipid-enveloped virus that causeshemorrhagic fever with a high fatality rate. Viral protein 40 (VP40) is the major EBOV matrix protein and regulates viral budding from the plasma membrane. VP40 is a transformer/morpheein that can structurally rearrange its native homodimer into either a hexameric filament that facilitates viral budding or a RNA-binding octameric ring that regulates viral transcription. VP40 associates with plasma-membrane lipids such as phosphatidylserine (PS), and this association is critical to budding from the host cell. However, it is poorly understood how different VP40 structures interact with PS, what essential residues are involved in this association, and whether VP40 has true selectivity for PS among different glycerophospholipid headgroups. In this study, we used lipid-binding assays, MD simulations, and cellular imaging to investigate the molecular basis of VP40-PS interactions and to determine whether different VP40 structures (i.e. monomer, dimer, and octamer) can interact with PScontaining membranes. Results from quantitative analysis indicated that VP40 associates with PS vesicles via a cationic patch in the C-terminal domain (Lys-224, 225 and Lys-274, 275). Substitutions of these residues with alanine reduced PSvesicle binding by >40-fold and abrogated VP40 localization to the plasma membrane. Dimeric VP40 had 2-fold greater affinity for PS-containing membranes than the monomer, whereas binding of the VP40 octameric ring was reduced by nearly 10-fold. Taken together, these results suggest http://www.jbc.org/cgi

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Section: Discussionsupporting

confidence: 86%

Section: Discussionmentioning

confidence: 99%

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A cationic, C-terminal patch and structural rearrangements in Ebola virus matrix VP40 protein control its interactions with phosphatidylserine

Vecchio

Frick

Jeevan

et al. 2018

Journal of Biological Chemistry

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134

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“…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). Additionally, treatment of human and mammalian cell lines expressing eVP40 with sphingosine did not have a significant effect on eVP40 plasma membrane localization (59,60).…”

Section: Discussionmentioning

confidence: 93%

“…Sphingosine, which carries a single positive charge, has been shown previously to neutralize the anionic charge of the plasma membrane (27). Delivery of sphingosine induced a significant loss of plasma membrane localization of proteins that interact with the plasma membrane based upon the anionic charge density but not of those proteins that specifically coordinate a lipid head group, such as PS or PIP 2 , for cellular localization (27,60). We delivered sphingosine to the cells expressing EGFP-MARV VP40 in an attempt to neutralize the plasma membrane anionic charge.…”

Section: Assessment Of Mvp40 Binding To Lipid Vesicles That Containedmentioning

confidence: 99%

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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Interdomain salt‐bridges in the Ebola virus protein VP40 and their role in domain association and plasma membrane localization

et al. 2016

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The Ebola virus protein VP40 is a transformer protein that possesses an extraordinary ability to accomplish multiple functions by transforming into various oligomeric conformations. The disengagement of the C-terminal domain (CTD) from the N-terminal domain (NTD) is a crucial step in the conformational transformations of VP40 from the dimeric form to the hexameric form or octameric ring structure. Here, we use various molecular dynamics (MD) simulations to investigate the dynamics of the VP40 protein and the roles of interdomain interactions that are important for the domain-domain association and dissociation, and report on experimental results of the behavior of mutant variants of VP40. The MD studies find that various salt-bridge interactions modulate the VP40 domain dynamics by providing conformational specificity through interdomain interactions. The MD simulations reveal a novel salt-bridge between D45-K326 when the CTD participates in a latch-like interaction with the NTD. The D45-K326 salt-bridge interaction is proposed to help domain-domain association, whereas the E76-K291 interaction is important for stabilizing the closed-form structure. The effects of the removal of important VP40 salt-bridges on plasma membrane (PM) localization, VP40 oligomerization, and virus like particle (VLP) budding assays were investigated experimentally by live cell imaging using an EGFP-tagged VP40 system. It is found that the mutations K291E and D45K show enhanced PM localization but D45K significantly reduced VLP formation.

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The Ebola Virus matrix protein, VP40, requires phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2) for extensive oligomerization at the plasma membrane and viral egress

Cited by 79 publications

References 62 publications

A cationic, C-terminal patch and structural rearrangements in Ebola virus matrix VP40 protein control its interactions with phosphatidylserine

A cationic, C-terminal patch and structural rearrangements in Ebola virus matrix VP40 protein control its interactions with phosphatidylserine

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

Interdomain salt‐bridges in the Ebola virus protein VP40 and their role in domain association and plasma membrane localization

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