Abstract

Enveloped viruses utilize fusion (F) proteins studding the surface of the virus to facilitate membrane fusion with a target cell membrane. Fusion of the viral envelope with a cellular membrane is required for release of viral genomic material, so the virus can ultimately reproduce and spread. To drive fusion, the F protein undergoes an irreversible conformational change, transitioning from a metastable pre-fusion conformation to a more thermodynamically stable post-fusion structure. Understanding the elements that control stability of the pre-fusion state and triggering to the post-fusion conformation is important for understanding F protein function. Mutations in F protein transmembrane (TM) domains implicated the TM domain in the fusion process, but the structural and molecular details in fusion remain unclear. Previously, analytical ultracentrifugation was utilized to demonstrate that isolated TM domains of Hendra virus F protein associate in a monomer-trimer equilibrium (Smith, E. C., Smith, S. E., Carter, J. R., Webb, S. R., Gibson, K. M., Hellman, L. M., Fried, M. G., and Dutch, R. E. (2013) J. Biol. Chem. 288, 35726–35735). To determine factors driving this association, 140 paramyxovirus F protein TM domain sequences were analyzed. A heptad repeat of β-branched residues was found, and analysis of the Hendra virus F TM domain revealed a heptad repeat leucine-isoleucine zipper motif (LIZ). Replacement of the LIZ with alanine resulted in dramatically reduced TM-TM association. Mutation of the LIZ in the whole protein resulted in decreased protein stability, including pre-fusion conformation stability. Together, our data suggest that the heptad repeat LIZ contributed to TM-TM association and is important for F protein function and pre-fusion stability.

Document Type

Article

Publication Date

2-17-2017

Notes/Citation Information

Published in The Journal of Biological Chemistry, v. 292, no. 14, p. 5685-5694.

This research was originally published in The Journal of Biological Chemistry. Stacy Webb, Tamas Nagy, Hunter Moseley, Michael Fried, and Rebecca Dutch. Hendra Virus Fusion Protein Transmembrane Domain Contributes to Pre-Fusion Protein Stability. J. Biol. Chem. 2017; 292:5685-5694. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

The copyright holder has granted the permission for posting the article here.

Digital Object Identifier (DOI)

https://doi.org/10.1074/jbc.M117.777235

Funding Information

This work was supported by National Institutes of Health Grants R01 AI051517, F31AI120653, and P30GM110787.

Related Content

This article contains supplemental Tables S1 and S2 and Figs. S1 and S2.

jbc.M117.777235-1.docx (149 kB)
Supplemental Data Figures 1 and 2

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