A model of the EIAV CA-SP hexamer (C6) from Gag-deltaMA spheres assembled at pH6

Structures of immature EIAV Gag lattices reveal a conserved role for IP6 in lentivirus assembly.

Rigid body fitting was done in Chimera. Missing residues were built de novo in Coot. Refinement was performed iteratively in Phenix and Coot.

CTF-correction was performed using NOVACTF

Gag construct was expressed in E.coli and purified using the SUMO-tag system.

Assembly was performed at pH6.

Tilt series were low-pass filtered according to their cumulative dose using exposure filters that were calculated using an exposure-dependent amplitude attenuation function and critical exposure constants (as published in Grant & Grigorieff, Elife, 2015). Tilt series were aligned and reconstructed in IMOD.

Data was acquired using a dose-symmetric tilt acquisition scheme, as described in Hagen et al, 2017, J. Struct. Biol, 197(2):191-8

Chimera was used to perform a rigid body fitting of one monomer of 2EIA into the electron microscopy density

The last residues of CA and the first residues of SP (T347-L359) were manually built in Coot

Virus-like-particles (spherical) of EIAV Gag deltaMAdeltap9 (referred to as Gag deltaMA) assembled at pH6.

Subtomogram extraction positions were defined in Amira using the electron microscopy toolbox by determing the radii and the center of the VLPs. Initially, positions were oversampled and subsequently cleaned during alignments using cross-correlation and distance thresholds.
Subvolumes were defined according to their position on the VLPs

MGDPLTWSKALKKLEKVTVQGSQKLTTGNCNWALSLVDLFHDTNFVKEKDWQLRDVIPLLEDVTQTLSGQEREAFERTWW
AISAVKMGLQINNVVDGKASFQLLRAKYEKKTANKKQSEPSEEYPIMIDGAGNRNFRPLTPRGYTTWVNTIQTNGLLNEA
SQNLFGILSVDCTSEEMNAFLDVVPGQAGQKQILLDAIDKIADDWDNRHPLPNAPLVAPPQGPIPMTARFIRGLGVPRER
QMEPAFDQFRQTYRQWIIEAMSEGIKVMIGKPKAQNIRQGAKEPYPEFVDRLLSQIKSEGHPQEISKFLTDTLTIQNANE
ECRNAMRHLRPEDTLEEKMYACRDIGTTKQKMMLLAKALQTGLAGPFKGGALKGGPLKAAQTCYNCGKPGHLSSQCRAPK
VCFKCKQPGHFSKQCRSVPKNGKQGAQGRPQKQTFPIQQKSQHNKSVVQETPQTQNLYPDLSEIKKEYNVKEKDQVEDLN
LDSLWE

National Institutes of Health/National Institute Of Allergy and Infectious Diseases (NIH/NIAID)

National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)

National Science Foundation (United States)

Medical Research Council (United Kingdom)

This entry contains the structure for EIAV Gag-deltaMA tubes assembled at pH8

This entry contains the structure for EIAV Gag-deltaMA spheres assembled at pH8

This entry contains the structure for EIAV Gag-deltaMA tubes assembled at pH6

This entry contains an example tomogram of EIAV Gag-deltaMA tubes and spheres assembled at pH6

Retrovirus, lentivirus, Equine infectious anemia virus, EIAV, Gag, capsid, IP6, phytic acid, inositolhexakiphosphate, VIRAL PROTEIN