Crystal structures of beta- and gammaretrovirus fusion proteins reveal a role for electrostatic stapling in viral entry

J Virol. 2014 Jan;88(1):143-53. doi: 10.1128/JVI.02023-13. Epub 2013 Oct 16.

Abstract

Membrane fusion is a key step in the life cycle of all envelope viruses, but this process is energetically unfavorable; the transmembrane fusion subunit (TM) of the virion-attached glycoprotein actively catalyzes the membrane merger process. Retroviral glycoproteins are the prototypical system to study pH-independent viral entry. In this study, we determined crystal structures of extramembrane regions of the TMs from Mason-Pfizer monkey virus (MPMV) and xenotropic murine leukemia virus-related virus (XMRV) at 1.7-Å and 2.2-Å resolution, respectively. The structures are comprised of a trimer of hairpins that is characteristic of class I viral fusion proteins and now completes a structural library of retroviral fusion proteins. Our results allowed us to identify a series of intra- and interchain electrostatic interactions in the heptad repeat and chain reversal regions. Mutagenesis reveals that charge-neutralizing salt bridge mutations significantly destabilize the postfusion six-helix bundle and abrogate retroviral infection, demonstrating that electrostatic stapling of the fusion subunit is essential for viral entry. Our data indicate that salt bridges are a major stabilizing force on the MPMV and XMRV retroviral TMs and likely provide the key energetics for viral and host membrane fusion.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Betaretrovirus / chemistry*
  • Betaretrovirus / physiology
  • Circular Dichroism
  • Crystallization
  • Gammaretrovirus / chemistry*
  • Gammaretrovirus / physiology
  • HEK293 Cells
  • Humans
  • Membrane Fusion*
  • Models, Molecular
  • Molecular Sequence Data
  • Protein Conformation
  • Recombinant Fusion Proteins / chemistry*
  • Sequence Homology, Amino Acid
  • Static Electricity*
  • Viral Envelope Proteins / chemistry*

Substances

  • Recombinant Fusion Proteins
  • Viral Envelope Proteins