Non-Enveloped Virus Entry: Structural Determinants and Mechanism of Functioning of a Viral Lytic Peptide.
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.
Kusuma School of Biological Sciences, Indian Institute of Technology-Delhi, Hauz Khas, New Delhi -110016, India.
Saumya
S
Bajaj
Kusuma School of Biological Sciences, Indian Institute of Technology-Delhi, Hauz Khas, New Delhi -110016, India.
Debajit
D
Dey
Kusuma School of Biological Sciences, Indian Institute of Technology-Delhi, Hauz Khas, New Delhi -110016, India.
Rohan
R
Bhukar
Kusuma School of Biological Sciences, Indian Institute of Technology-Delhi, Hauz Khas, New Delhi -110016, India.
Mohit
M
Kumar
Kusuma School of Biological Sciences, Indian Institute of Technology-Delhi, Hauz Khas, New Delhi -110016, India. Electronic address: mbanerjee@bioschool.iitd.ac.in.
Manidipa
M
Banerjee
D000818:Animals; D055592:Biophysical Phenomena; D002460:Cell Line; D002462:Cell Membrane / Q000821:virology; D020285:Cryoelectron Microscopy; D007313:Insecta; D008954:Models, Biological; D008958:Models, Molecular; D056004:Molecular Dynamics Simulation; D030381:Nodaviridae / Q000502:physiology*; D011487:Protein Conformation; D055503:Protein Multimerization; D014764:Viral Proteins / Q000737:chemistry* / Q000378:metabolism*; D053586:Virus Internalization
D014764:Viral Proteins
cryo-electron microscopy; membrane lytic peptide; molecular dynamics simulation; non-enveloped virus; virus entry