Non-Enveloped Virus Entry: Structural Determinants and Mechanism of Functioning of a Viral Lytic Peptide

Saumya Bajaj; Debajit Dey; Rohan Bhukar; Mohit Kumar; Manidipa Banerjee

doi:10.1016/j.jmb.2016.06.006

Non-Enveloped Virus Entry: Structural Determinants and Mechanism of Functioning of a Viral Lytic Peptide

Saumya Bajaj, Debajit Dey, Rohan Bhukar, Mohit Kumar, Manidipa Banerjee

Biology

Research output: Contribution to journal › Article › peer-review

Abstract

In the absence of lipid envelopes and associated fusion proteins, non-enveloped viruses employ membrane lytic peptides to breach the limiting membranes of host cells. Although several of these lytic peptides have been identified and characterized, their manner of deployment and interaction with host membranes remains unclear in most cases. We are using the gamma peptide of Flock House Virus (FHV), a model non-enveloped virus, to understand the mechanistic details of non-enveloped virus interaction with host cell membranes. We utilized a combination of biophysical assays, molecular dynamics simulation studies, and single-particle cryo-electron microscopy to elucidate the functional and structural determinants for membrane penetration by gamma in context of the FHV capsid. Although the amphipathic, helical N-terminal region of gamma (γ1) was previously thought to be the membrane-penetrating module, with the C-terminal region having a supporting role in correct structural positioning of γ1, we demonstrate that the C terminus of gamma directly participates in membrane penetration. Our studies suggest that full-length gamma, including the hydrophobic C terminus, forms an alpha-helical hairpin motif, and any disruption in this motif drastically reduces its functionality, in spite of the correct positioning of amphipathic γ1 in the virus capsid. Taken together, our data suggest that the most effective module for membrane disruption is a pentameric unit of full-length gamma, released from the virus, which associates with membranes via both N- and C-terminal ends.

Original language	English
Pages (from-to)	3540-56
Number of pages	17
Journal	Journal of Molecular Biology
Volume	428
Issue number	17
DOIs	https://doi.org/10.1016/j.jmb.2016.06.006
Publication status	Published - 28 Aug 2016

Bibliographical note

Keywords

Animals
Biophysical Phenomena
Cell Line
Cell Membrane/virology
Cryoelectron Microscopy
Insecta
Models, Biological
Models, Molecular
Molecular Dynamics Simulation
Nodaviridae/physiology
Protein Conformation
Protein Multimerization
Viral Proteins/chemistry
Virus Internalization

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10.1016/j.jmb.2016.06.006

Cite this

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title = "Non-Enveloped Virus Entry: Structural Determinants and Mechanism of Functioning of a Viral Lytic Peptide",

abstract = "In the absence of lipid envelopes and associated fusion proteins, non-enveloped viruses employ membrane lytic peptides to breach the limiting membranes of host cells. Although several of these lytic peptides have been identified and characterized, their manner of deployment and interaction with host membranes remains unclear in most cases. We are using the gamma peptide of Flock House Virus (FHV), a model non-enveloped virus, to understand the mechanistic details of non-enveloped virus interaction with host cell membranes. We utilized a combination of biophysical assays, molecular dynamics simulation studies, and single-particle cryo-electron microscopy to elucidate the functional and structural determinants for membrane penetration by gamma in context of the FHV capsid. Although the amphipathic, helical N-terminal region of gamma (γ1) was previously thought to be the membrane-penetrating module, with the C-terminal region having a supporting role in correct structural positioning of γ1, we demonstrate that the C terminus of gamma directly participates in membrane penetration. Our studies suggest that full-length gamma, including the hydrophobic C terminus, forms an alpha-helical hairpin motif, and any disruption in this motif drastically reduces its functionality, in spite of the correct positioning of amphipathic γ1 in the virus capsid. Taken together, our data suggest that the most effective module for membrane disruption is a pentameric unit of full-length gamma, released from the virus, which associates with membranes via both N- and C-terminal ends.",

keywords = "Animals, Biophysical Phenomena, Cell Line, Cell Membrane/virology, Cryoelectron Microscopy, Insecta, Models, Biological, Models, Molecular, Molecular Dynamics Simulation, Nodaviridae/physiology, Protein Conformation, Protein Multimerization, Viral Proteins/chemistry, Virus Internalization",

author = "Saumya Bajaj and Debajit Dey and Rohan Bhukar and Mohit Kumar and Manidipa Banerjee",

year = "2016",

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T2 - Structural Determinants and Mechanism of Functioning of a Viral Lytic Peptide

AU - Bajaj, Saumya

AU - Dey, Debajit

AU - Bhukar, Rohan

AU - Kumar, Mohit

AU - Banerjee, Manidipa

PY - 2016/8/28

Y1 - 2016/8/28

N2 - In the absence of lipid envelopes and associated fusion proteins, non-enveloped viruses employ membrane lytic peptides to breach the limiting membranes of host cells. Although several of these lytic peptides have been identified and characterized, their manner of deployment and interaction with host membranes remains unclear in most cases. We are using the gamma peptide of Flock House Virus (FHV), a model non-enveloped virus, to understand the mechanistic details of non-enveloped virus interaction with host cell membranes. We utilized a combination of biophysical assays, molecular dynamics simulation studies, and single-particle cryo-electron microscopy to elucidate the functional and structural determinants for membrane penetration by gamma in context of the FHV capsid. Although the amphipathic, helical N-terminal region of gamma (γ1) was previously thought to be the membrane-penetrating module, with the C-terminal region having a supporting role in correct structural positioning of γ1, we demonstrate that the C terminus of gamma directly participates in membrane penetration. Our studies suggest that full-length gamma, including the hydrophobic C terminus, forms an alpha-helical hairpin motif, and any disruption in this motif drastically reduces its functionality, in spite of the correct positioning of amphipathic γ1 in the virus capsid. Taken together, our data suggest that the most effective module for membrane disruption is a pentameric unit of full-length gamma, released from the virus, which associates with membranes via both N- and C-terminal ends.

AB - In the absence of lipid envelopes and associated fusion proteins, non-enveloped viruses employ membrane lytic peptides to breach the limiting membranes of host cells. Although several of these lytic peptides have been identified and characterized, their manner of deployment and interaction with host membranes remains unclear in most cases. We are using the gamma peptide of Flock House Virus (FHV), a model non-enveloped virus, to understand the mechanistic details of non-enveloped virus interaction with host cell membranes. We utilized a combination of biophysical assays, molecular dynamics simulation studies, and single-particle cryo-electron microscopy to elucidate the functional and structural determinants for membrane penetration by gamma in context of the FHV capsid. Although the amphipathic, helical N-terminal region of gamma (γ1) was previously thought to be the membrane-penetrating module, with the C-terminal region having a supporting role in correct structural positioning of γ1, we demonstrate that the C terminus of gamma directly participates in membrane penetration. Our studies suggest that full-length gamma, including the hydrophobic C terminus, forms an alpha-helical hairpin motif, and any disruption in this motif drastically reduces its functionality, in spite of the correct positioning of amphipathic γ1 in the virus capsid. Taken together, our data suggest that the most effective module for membrane disruption is a pentameric unit of full-length gamma, released from the virus, which associates with membranes via both N- and C-terminal ends.

KW - Animals

KW - Biophysical Phenomena

KW - Cell Line

KW - Cell Membrane/virology

KW - Cryoelectron Microscopy

KW - Insecta

KW - Models, Biological

KW - Models, Molecular

KW - Molecular Dynamics Simulation

KW - Nodaviridae/physiology

KW - Protein Conformation

KW - Protein Multimerization

KW - Viral Proteins/chemistry

KW - Virus Internalization

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DO - 10.1016/j.jmb.2016.06.006

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