National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)
124279
United States
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)
118047
United States
National Science Foundation (NSF, United States)
DMR-2011401
United States
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)
124165
United States
National Institutes of Health/National Institute on Drug Abuse (NIH/NIDA)
007097
United States
National Institutes of Health/National Institute Of Allergy and Infectious Diseases (NIH/NIAID)
150351
United States
National Institutes of Health/National Institute Of Allergy and Infectious Diseases (NIH/NIAID)
83196
United States
Citation
Journal: J Mol Biol / Year: 2023 Title: HIV-2 Immature Particle Morphology Provides Insights into Gag Lattice Stability and Virus Maturation. Authors: Nathaniel Talledge / Huixin Yang / Ke Shi / Raffaele Coray / Guichuan Yu / William G Arndt / Shuyu Meng / Gloria C Baxter / Luiza M Mendonça / Daniel Castaño-Díez / Hideki Aihara / Louis ...Authors: Nathaniel Talledge / Huixin Yang / Ke Shi / Raffaele Coray / Guichuan Yu / William G Arndt / Shuyu Meng / Gloria C Baxter / Luiza M Mendonça / Daniel Castaño-Díez / Hideki Aihara / Louis M Mansky / Wei Zhang / Abstract: Retrovirus immature particle morphology consists of a membrane enclosed, pleomorphic, spherical and incomplete lattice of Gag hexamers. Previously, we demonstrated that human immunodeficiency virus ...Retrovirus immature particle morphology consists of a membrane enclosed, pleomorphic, spherical and incomplete lattice of Gag hexamers. Previously, we demonstrated that human immunodeficiency virus type 2 (HIV-2) immature particles possess a distinct and extensive Gag lattice morphology. To better understand the nature of the continuously curved hexagonal Gag lattice, we have used the single particle cryo-electron microscopy method to determine the HIV-2 Gag lattice structure for immature virions. The reconstruction map at 5.5 Å resolution revealed a stable, wineglass-shaped Gag hexamer structure with structural features consistent with other lentiviral immature Gag lattice structures. Cryo-electron tomography provided evidence for nearly complete ordered Gag lattice structures in HIV-2 immature particles. We also solved a 1.98 Å resolution crystal structure of the carboxyl-terminal domain (CTD) of the HIV-2 capsid (CA) protein that identified a structured helix 12 supported via an interaction of helix 10 in the absence of the SP1 region of Gag. Residues at the helix 10-12 interface proved critical in maintaining HIV-2 particle release and infectivity. Taken together, our findings provide the first 3D organization of HIV-2 immature Gag lattice and important insights into both HIV Gag lattice stabilization and virus maturation.
Details: 100 mM NaCl, 10 mM Tris-HCL pH 8.0, 1mM EDTA
Grid
Model: Quantifoil R2/2 / Material: COPPER / Mesh: 200 / Support film - Material: CARBON / Support film - topology: HOLEY / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 30 sec. / Pretreatment - Atmosphere: AIR / Details: Leica ACE600 Glow Discharger.
Vitrification
Cryogen name: ETHANE / Chamber humidity: 85 % / Chamber temperature: 292.15 K / Instrument: LEICA EM GP / Details: Leica GP2 Grid Plunger..
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Electron microscopy
Microscope
FEI TITAN KRIOS
Specialist optics
Energy filter - Slit width: 20 eV
Image recording
Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: SUPER-RESOLUTION / Digitization - Frames/image: 1-40 / Number grids imaged: 10 / Number real images: 2508 / Average exposure time: 10.0 sec. / Average electron dose: 50.0 e/Å2
Electron beam
Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
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