Ebola and Marburg virus matrix layers are locally ordered assemblies of VP40 dimers

Wan, William; Clarke, Mairi; Norris, Michael; Kolesnikova, Larissa; Koehler, Alexander; Bornholdt, Zachary A.; Becker, Stephan; Saphire, Erica Ollmann; Briggs, John

doi:10.7554/elife.59225

Cited by 51 publications

(93 citation statements)

References 48 publications

(91 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…VP40 connects the viral nucleocapsid to the inside of the cell membrane and promotes the egress event to achieve the whole viral life cycle, which depend on its oligomerisation ability [ 13 ]. When the C-terminal domain (CTD) of VP40 attaches to cell membranes [ 14 ], it then forms polymers through self-interactions of N-terminal domain (NTD), which contributes to the release of EBOV particles [ 15 ]. VP40 can form linear hexamers and octamer rings [ 16 ]: the hexameric VP40 plays a great role in lipid raft and viral particle formation [ 17 , 18 ]; the octameric VP40 is essential for binding and viral life cycle [ 8 , 19 ].…”

Section: Introductionmentioning

confidence: 99%

P300-mediated NEDD4 acetylation drives ebolavirus VP40 egress by enhancing NEDD4 ligase activity

et al. 2021

View full text Add to dashboard Cite

The final stage of Ebola virus (EBOV) replication is budding from host cells, where the matrix protein VP40 is essential for driving this process. Many post-translational modifications such as ubiquitination are involved in VP40 egress, but acetylation has not been studied yet. Here, we characterize NEDD4 is acetylated at a conserved Lys667 mediated by the acetyltransferase P300 which drives VP40 egress process. Importantly, P300-mediated NEDD4 acetylation promotes NEDD4-VP40 interaction which enhances NEDD4 E3 ligase activity and is essential for the activation of VP40 ubiquitination and subsequent egress. Finally, we find that Zaire ebolavirus production is dramatically reduced in P300 knockout cell lines, suggesting that P300-mediated NEDD4 acetylation may have a physiological effect on Ebola virus life cycle. Thus, our study identifies an acetylation-dependent regulatory mechanism that governs VP40 ubiquitination and provides insights into how acetylation controls EBOV VP40 egress.

show abstract

Section: Introductionmentioning

confidence: 99%

P300-mediated NEDD4 acetylation drives ebolavirus VP40 egress by enhancing NEDD4 ligase activity

et al. 2021

View full text Add to dashboard Cite

show abstract

“…The hydrophobic interaction at the hexamer-hexamer interface may provide an agile interface, giving flexibility to the filaments. (E) Zoom into hexamer-hexamer interface in the mVP40 filament formed through CTD-CTD linear oligomerization as proposed by Wan et al (Wan et al , 2020). (CTD from each monomer is in showed in different colors)…”

Section: Resultsmentioning

confidence: 94%

“…It is possible that NTD-NTD interactions are required to increase membrane bending, elongation of tubule and/or for host cell factor recruitment at assembly sites. Furthermore, using the recent model proposed in CTD-CTD linear oligomerization ((Wan et al , 2020), Fig. S6), we simulated the CTD-CTD complex that indicated this interface may involve Met 191 , Asn 222 , Tyr 195 and Leu 226 as we report here (Fig.…”

Section: Discussionmentioning

confidence: 92%

“…CTD-CTD-interactions may be required at a specific stage of the matrix assembly after protein associates with the plasma membrane and establishment of NTD-NTD interactions to initiate the protein high ordered-oligomerization. A recent model proposed CTD-CTD linear oligomerization that is most likely in both MARV and EBOV virions and VLPs (Wan et al , 2020). However, our study suggests the importance of NTD-NTD oligomerization in cells and in vitro to establish the building blocks for higher-ordered oligomer formation and particle release.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Lipid-specific protein oligomerization is regulated by two interfaces in Marburg virus matrix protein VP40

Amiar

et al. 2020

Preprint

View full text Add to dashboard Cite

SummaryMarburg virus major matrix protein (mVP40) dimers associate with anionic lipids at the plasma membrane and undergo a dynamic and extensive self-oligomerization into the structural matrix layer which confers the virion shape and stability. Using a myriad of in vitro and cellular techniques, we present a mVP40 assembly model highlighting two distinct oligomerization interfaces (N-terminal domain (NTD) and C-terminal domain (CTD)) in mVP40. Cellular studies of NTD and CTD oligomerization interface mutants demonstrated the importance of each interface in the mVP40 matrix assembly through protein trafficking to the plasma membrane and homo-multimerization that induced protein enrichment, plasma membrane fluidity changes and elongations at the plasma membrane. A novel APEX-TEM method was employed to closely assess the ultrastructural localization of and formation of viral particles for wild type and mutants. Taken together, these studies present a mechanistic model of mVP40 oligomerization and assembly at the plasma membrane during virion assembly.

show abstract

“…VP40 is sufficient to drive virion budding and is responsible for the characteristic filamentous shape [23]. Recent cryo-ET studies revealed the organization of VP40 proteins in budded particles into an apparent helical scaffold lining the inner viral membrane leaflet [21]. Besides VP40 as a major driver of EBOV budding, actin has been shown to be indispensable, since the disruption of the actin cytoskeleton abrogates particle formation [24].…”

Section: Introductionmentioning

confidence: 99%

Dual-axis Volta phase plate cryo-electron tomography of Ebola virus-like particles reveals actin-VP40 interactions

Winter

Chlanda

2021

Preprint

View full text Add to dashboard Cite

Cryo-electron tomography (cryo-ET) is a pivotal imaging technique for studying the structure of pleomorphic enveloped viruses and their interactions with the host at native conditions. Owing to the limited tilting range of samples with a slab geometry, electron tomograms suffer from so-called missing wedge information in Fourier space. In dual-axis cryo-ET, two tomograms reconstructed from orthogonally oriented tilt series are combined into a tomogram with improved resolution as the missing wedge information is reduced to a pyramid. Volta phase plate (VPP) allows to perform in-focus cryo-ET with high contrast transfer at low-resolution frequencies and thus its application may improve the quality of dual-axis tomograms. Here, we compare dual-axis cryo-ET with and without VPP on Ebola virus-like particles to visualize and segment viral and host cell proteins within the membrane-enveloped filamentous particles. Dual-axis VPP cryo-ET reduces the missing wedge information and ray artifacts arising from the weighted back-projection during tomogram reconstruction, thereby minimizing ambiguity in the analysis of crowded environments and facilitating 3D segmentation. We show that dual-axis VPP tomograms provide a comprehensive description of macromolecular organizations such as nucleocapsid assembly states, the distribution of glycoproteins on the viral envelope and asymmetric arrangements of the VP40 layer in non-filamentous regions of virus-like particles. Our data reveal actin filaments within virus-like particles in close proximity to the viral VP40 scaffold, suggesting a direct interaction between VP40 and actin filaments. Dual-axis VPP cryo-ET provides more complete 3D information at high contrast and allows for better interpretation of macromolecule interactions and pleomorphic organizations.

show abstract

Ebola and Marburg virus matrix layers are locally ordered assemblies of VP40 dimers

Cited by 51 publications

References 48 publications

P300-mediated NEDD4 acetylation drives ebolavirus VP40 egress by enhancing NEDD4 ligase activity

P300-mediated NEDD4 acetylation drives ebolavirus VP40 egress by enhancing NEDD4 ligase activity

Lipid-specific protein oligomerization is regulated by two interfaces in Marburg virus matrix protein VP40

Dual-axis Volta phase plate cryo-electron tomography of Ebola virus-like particles reveals actin-VP40 interactions

Contact Info

Product

Resources

About