National Institutes of Health/National Institute Of Allergy and Infectious Diseases (NIH/NIAID)
R01GM151775
United States
National Institutes of Health/National Institute Of Allergy and Infectious Diseases (NIH/NIAID)
R21DE032878
United States
National Institutes of Health/National Institute Of Allergy and Infectious Diseases (NIH/NIAID)
F31AI184673
United States
National Institutes of Health/National Institute Of Allergy and Infectious Diseases (NIH/NIAID)
T32AI083196
United States
Citation
Journal: Nat Commun / Year: 2025 Title: High-resolution analysis of the human T-cell leukemia virus capsid protein reveals insights into immature particle morphology. Authors: William G Arndt / Alireza Ramezani / Nathaniel Talledge / Guichuan Yu / Huixin Yang / Bo Chen / Juan R Perilla / Wei Zhang / Louis M Mansky / Abstract: Infection with human T-cell leukemia virus type 1 (HTLV-1) can result in adult T-cell leukemia/lymphoma and HTLV-1 associated-myelopathy/tropical spastic paraparesis. The Gag polyprotein - the major ...Infection with human T-cell leukemia virus type 1 (HTLV-1) can result in adult T-cell leukemia/lymphoma and HTLV-1 associated-myelopathy/tropical spastic paraparesis. The Gag polyprotein - the major structural protein - is crucial for driving virus particle assembly, with the capsid (CA) domain as the key determinant for Gag multimerization. Here, we characterize the immature CA lattice from immature virus particles by using cryo-electron microscopy and tomography (cryo-EM/ET). We report resolving the immature CA lattice to 3.4 Å resolution by single particle analysis (SPA). Our reconstruction reveals that the lattice is stabilized through a trimeric NTD inter-hexamer interface and a dimeric CTD inter-hexamer interface. Further analysis by cryo-ET reveals clear heterogeneity, notably the varying lattice curvatures and the varying distances from the CA layer to the membrane. Intriguingly, inositol hexakisphosphate (IP6) is dispensable for HTLV-1 immature particle assembly and proper immature lattice formation. These observations provide deeper insights into the molecular basis of HTLV-1 immature particle morphology as well as aid in revealing therapeutic targets.
Supramolecule #1: Human T-cell leukemia virus type I
Supramolecule
Name: Human T-cell leukemia virus type I / type: virus / ID: 1 / Parent: 0 / Macromolecule list: all Details: HTLV-1 virus-like particles produced by transfecting 293T cells with HTLV-1 Gag and Envelope. NCBI-ID: 11908 / Sci species name: Human T-cell leukemia virus type I / Sci species strain: B1033-2009 / Virus type: VIRUS-LIKE PARTICLE / Virus isolate: STRAIN / Virus enveloped: Yes / Virus empty: No
Host (natural)
Organism: Homo sapiens (human)
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Macromolecule #1: Gag protein
Macromolecule
Name: Gag protein / type: protein_or_peptide / ID: 1 / Number of copies: 3 / Enantiomer: LEVO
Source (natural)
Organism: Human T-cell leukemia virus type I / Strain: B1033-2009
pH: 8 / Details: 100 mM NaCl, 10 mM Tris-HCL, 1 mM EDTA
Grid
Model: Quantifoil R2/1 / Material: COPPER / Mesh: 200 / Support film - Material: CARBON / Support film - topology: HOLEY / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 30 sec. / Pretreatment - Atmosphere: AIR
Vitrification
Cryogen name: ETHANE / Instrument: LEICA EM GP / Details: Leica GP2 Grid Plunger.
Details
The virus particles were purified from the cell culture supernatant via optiprep gradient ultracentrifugation.
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Electron microscopy
Microscope
TFS 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: 1 / Number real images: 2502 / Average electron dose: 50.0 e/Å2
Electron beam
Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
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