Open AccessCryo-electron Microscopy Structure of the Native Prototype Foamy Virus Glycoprotein and Virus Architecture
Author(s) -
Grégory Effantin,
Leandro F. Estrozi,
Nick Aschman,
Patricia Renesto,
Nicole Stanke,
Dirk Lindemann,
Guy Schoehn,
Winfríed Weissenhorn
Publication year - 2016
Publication title -
plos pathogens
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.719
H-Index - 206
eISSN - 1553-7374
pISSN - 1553-7366
DOI - 10.1371/journal.ppat.1005721
Subject(s) - cryo electron microscopy , capsid , trimer , protein subunit , transmembrane protein , biophysics , transmembrane domain , lipid bilayer fusion , glycoprotein , biology , cryo electron tomography , virus , cytoplasm , chemistry , crystallography , microbiology and biotechnology , virology , membrane , gene , biochemistry , physics , receptor , dimer , organic chemistry , optics , tomography
Foamy viruses (FV) belong to the genus Spumavirus, which forms a distinct lineage in the Retroviridae family. Although the infection in natural hosts and zoonotic transmission to humans is asymptomatic, FVs can replicate well in human cells making it an attractive gene therapy vector candidate. Here we present cryo-electron microscopy and (cryo-)electron tomography ultrastructural data on purified prototype FV (PFV) and PFV infected cells. Mature PFV particles have a distinct morphology with a capsid of constant dimension as well as a less ordered shell of density between the capsid and the membrane likely formed by the Gag N-terminal domain and the cytoplasmic part of the Env leader peptide gp18 LP . The viral membrane contains trimeric Env glycoproteins partly arranged in interlocked hexagonal assemblies. In situ 3D reconstruction by subtomogram averaging of wild type Env and of a Env gp48 TM - gp80 SU cleavage site mutant showed a similar spike architecture as well as stabilization of the hexagonal lattice by clear connections between lower densities of neighboring trimers. Cryo-EM was employed to obtain a 9 Å resolution map of the glycoprotein in its pre-fusion state, which revealed extensive trimer interactions by the receptor binding subunit gp80 SU at the top of the spike and three central helices derived from the fusion protein subunit gp48 TM . The lower part of Env, presumably composed of interlaced parts of gp48 TM , gp80 SU and gp18 LP anchors the spike at the membrane. We propose that the gp48 TM density continues into three central transmembrane helices, which interact with three outer transmembrane helices derived from gp18 LP . Our ultrastructural data and 9 Å resolution glycoprotein structure provide important new insights into the molecular architecture of PFV and its distinct evolutionary relationship with other members of the Retroviridae .