A stochastic assembly model for Nipah virus revealed by super-resolution microscopy

Liu, Qian; Chen, Lei; Aguilar, Hector C.; Chou, Kuang-Chao

doi:10.1038/s41467-018-05480-2

Cited by 29 publications

(30 citation statements)

References 37 publications

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“…Work with the paramyxovirus MeV has shown that the F protein is enriched in lipid rafts, and this partitioning is important for MeV assembly at the plasma membrane (49). Studies of NiV have shown evidence of F protein clustering in the plasma membrane, suggesting that membrane domains may be needed for proper surface glycoprotein organization for assembly and fusion (54). For Newcastle disease virus, lipid rafts have been shown to participate in forming and maintaining F protein and attachment protein complexes in the plasma membrane (55).…”

Section: Discussionmentioning

confidence: 99%

Transmembrane Domain Dissociation Is Required for Hendra Virus F Protein Fusogenic Activity

Slaughter

Dutch

2019

J Virol

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Hendra virus (HeV) is a zoonotic paramyxovirus that utilizes a trimeric fusion (F) protein within its lipid bilayer to mediate membrane merger with a cell membrane for entry. Previous HeV F studies showed that transmembrane domain (TMD) interactions are important for stabilizing the prefusion conformation of the protein prior to triggering. Thus, the current model for HeV F fusion suggests that modulation of TMD interactions is critical for initiation and completion of conformational changes that drive membrane fusion. HeV F constructs (T483C/V484C, V484C/N485C, and N485C/P486C) were generated with double cysteine substitutions near the N-terminal region of the TMD to study the effect of altered flexibility in this region. Oligomeric analysis showed that the double cysteine substitutions successfully promoted intersubunit disulfide bond formation in HeV F. Subsequent fusion assays indicated that the introduction of disulfide bonds in the mutants prohibited fusion events. Further testing confirmed that T483C/V484C and V484C/N485C were expressed at the cell surface at levels that would allow for fusion. Attempts to restore fusion with a reducing agent were unsuccessful, suggesting that the introduced disulfide bonds were likely buried in the membrane. Conformational analysis showed that T483C/V484C and V484C/N485C were able to bind a prefusion conformation-specific antibody prior to cell disruption, indicating that the introduced disulfide bonds did not significantly affect protein folding. This study is the first to report that TMD dissociation is required for HeV F fusogenic activity and strengthens our model for HeV fusion. IMPORTANCE The paramyxovirus Hendra virus (HeV) causes severe respiratory illness and encephalitis in humans. To develop therapeutics for HeV and related viral infections, further studies are needed to understand the mechanisms underlying paramyxovirus fusion events. Knowledge gained in studies of the HeV fusion (F) protein may be applicable to a broad span of enveloped viruses. In this study, we demonstrate that disulfide bonds introduced between the HeV F transmembrane domains (TMDs) block fusion. Depending on the location of these disulfide bonds, HeV F can still fold properly and bind a prefusion conformation-specific antibody prior to cell disruption. These findings support our current model for HeV membrane fusion and expand our knowledge of the TMD and its role in HeV F stability and fusion promotion.

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Section: Discussionmentioning

confidence: 99%

Transmembrane Domain Dissociation Is Required for Hendra Virus F Protein Fusogenic Activity

Slaughter

Dutch

2019

J Virol

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“…Further analysis showed that the proper endocytosis and recycling of the F protein, mediated by residues S490 and Y498, were critical for proper virus-like particle (VLP) formation [120]. This suggests that residues in the TMD participate in viral assembly by facilitating specific intracellular trafficking in Hendra and Nipah viruses, but the extent and mechanism remain unclear, as there are conflicting results on the nature of F trafficking and incorporation in Nipah VLPs [126,127]. Residues in the F protein ectodomain likely also assist in proper F protein incorporation into viral particles, since a chimera of the Rabies virus particle ectodomain and NDV TMD and CTD demonstrated inefficient incorporation into NDV viral particles [122].…”

Section: Paramyxovirusesmentioning

confidence: 99%

Viral Membrane Fusion and the Transmembrane Domain

Barrett

Dutch

2020

Viruses

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Initiation of host cell infection by an enveloped virus requires a viral-to-host cell membrane fusion event. This event is mediated by at least one viral transmembrane glycoprotein, termed the fusion protein, which is a key therapeutic target. Viral fusion proteins have been studied for decades, and numerous critical insights into their function have been elucidated. However, the transmembrane region remains one of the most poorly understood facets of these proteins. In the past ten years, the field has made significant advances in understanding the role of the membrane-spanning region of viral fusion proteins. We summarize developments made in the past decade that have contributed to the understanding of the transmembrane region of viral fusion proteins, highlighting not only their critical role in the membrane fusion process, but further demonstrating their involvement in several aspects of the viral lifecycle.

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“…A combination of PALM and STORM techniques has also been used to simultaneously image multiple components within the cell membrane. [64][65][66][67][68][69][70][71][72][73] "Ordered" and "disordered" lipid domains as well as the localization of clustered B cell receptor into ordered domains were measured in mouse B lymphoma cell membranes using this approach (Figure 1). 69 The combinations of fluorescent probes in this study were Atto 655 with Alexa Fluor 532 (i.e., dual Laser irradiances were adjusted between 5 to 20 kW/cm 2 to achieve favorable conditions for single molecule/protein localization, and the emission was separated into two channels prior to reaching the detector.…”

Section: Single Molecule Localization Microscopy (Smlm)mentioning

confidence: 99%

“…In another study utilizing PALM/STORM, the organization of Nipah virus proteins on the plasma membrane was detected. 73 Nipah is a biosafety level 4 human-to-human transmitted virus. The results showed clusters of virus proteins, such as attachment glycoproteins and fusion glycoproteins, were randomly distributed on the mammalian PK13 cell membrane regardless of whether virus matrix proteins were present or absent.…”

Section: Figurementioning

confidence: 99%

Optical Imaging of the Nanoscale Structure and Dynamics of Biological Membranes

2018

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Biological membranes serve as the fundamental unit of life, allowing the compartmentalization of cellular contents into subunits with specific functions. The bilayer structure, consisting of lipids, proteins, small molecules, and sugars, also serves many other complex functions in addition to maintaining the relative stability of the inner compartments. Signal transduction, regulation of solute exchange, active transport, and energy transduction through ion gradients all take place at biological membranes, primarily with the assistance of membrane proteins. For these functions, membrane structure is often critical. The fluidmosaic model introduced by Singer and Nicolson in 1972 evokes the dynamic and fluid nature of biological membranes.(1) According to this model, integral and peripheral proteins are oriented in a viscous phospholipid bilayer. Both proteins and lipids can diffuse laterally through the two-dimensional structure. Modern experimental evidence has shown, however, that the structure of the membrane is considerably more complex; various domains in the biological membranes, such as lipid rafts and confinement regions, form a more complicated molecular organization. The proper organization and dynamics of the membrane components are critical for the function of the entire cell. For example, cell signaling is often initiated at biological membranes and requires receptors to diffuse and assemble into complexes and clusters, and the resulting downstream events have consequences throughout the cell. Revealing the molecular level details of these signaling events is the foundation to understanding numerous unsolved questions regarding cellular life.

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A stochastic assembly model for Nipah virus revealed by super-resolution microscopy

Cited by 29 publications

References 37 publications

Transmembrane Domain Dissociation Is Required for Hendra Virus F Protein Fusogenic Activity

Transmembrane Domain Dissociation Is Required for Hendra Virus F Protein Fusogenic Activity

Viral Membrane Fusion and the Transmembrane Domain

Optical Imaging of the Nanoscale Structure and Dynamics of Biological Membranes

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