Human Immunodeficiency Virus Type 2 Capsid Protein Mutagenesis Reveals Amino Acid Residues Important for Virus Particle Assembly

Yang, Huixin; Talledge, Nathaniel; Arndt, William G.; Zhang, Wei; Mansky, Louis M.

doi:10.1016/j.jmb.2022.167753

Cited by 5 publications

(4 citation statements)

References 60 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…For example, at the Gag lattice three-fold interface, we found that exchanging HIV-1 and HIV-2 residues significantly reduced virus particle production, and diminished virus particle infectivity. Mutations of a non-conserved residue in helix 2 of the CA proteinsi.e., HIV-2 CA G38M and HIV-1 CA M39Greduced by 50% virus particle production as well as eliminating virus infectivity [54]. Additionally, the HIV-2 CA N127E mutant, which introduced the HIV-1 encoding residue into HIV-2 CA, severely reduced particle production.…”

Section: Discussionmentioning

confidence: 99%

HIV-2 Immature Particle Morphology Provides Insights into Gag Lattice Stability and Virus Maturation

Talledge

Yang

Shi

et al. 2023

Journal of Molecular Biology

Self Cite

View full text Add to dashboard Cite

Section: Discussionmentioning

confidence: 99%

HIV-2 Immature Particle Morphology Provides Insights into Gag Lattice Stability and Virus Maturation

Talledge

Yang

Shi

et al. 2023

Journal of Molecular Biology

Self Cite

View full text Add to dashboard Cite

“…For example, at the Gag lattice three-fold interface, we found that exchanging HIV-1 and HIV-2 residues significantly reduced virus particle production, and diminished virus particle infectivity. Mutations of a non-conserved residue in helix 2 of the CA proteins – i.e ., HIV-2 CA G38M and HIV-1 CA M39G – reduced by 50% virus particle production as well as eliminating virus infectivity [45]. Additionally, the HIV-2 CA N127E mutant, which introduced the HIV-1 encoding residue into HIV-2 CA, severely reduced particle production.…”

Section: Discussionmentioning

confidence: 99%

HIV-2 Immature Particle Morphology Provides Insights into Gag Lattice Stability and Virus Maturation

Talledge

Yang

Shi

et al. 2022

Preprint

View full text Add to dashboard Cite

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 used single particle cryo-electron microscopy to determine the HIV-2 Gag lattice structure for immature virions. The reconstruction map revealed a stable, wineglass-shaped Gag hexamer structure with structural features consistent with other lentiviral immature Gag structures. 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 evidence for a novel stabilization interface mediated by the HIV-2 CACTD that delivers important clues for HIV Gag lattice stabilization as well as important insights into virus maturation.

show abstract

“…The pN3-HTLV-1-Gag plasmid and the HTLV-1 Env expression plasmids were co-transfected into HEK293T/17 cells by using GenJet, ver II, at a 10:1 molar ratio as previously described [51, 52]. At 48-h post-transfection, cell culture supernatants were harvested and centrifuged at 1,800 × g for 10 min and followed by passing through a 0.2 µm filter.…”

Section: Methodsmentioning

confidence: 99%

“…Gag subcellular distribution was analyzed by quantifying the degree of Gag puncta formation in cells by using confocal laser scanning microscopy to localize Gag-eYFP as previously described [21]. Briefly, HeLa cells were cultured in six-well plates on 1.5 standard glass coverslips coated with poly-L-lysine as previously described [21, 52]. HeLa cells were transiently transfected with EYFP-tagged Gag plasmids by using GenJet, ver II (SignaGen, Gaithersburg, MD).…”

Section: Methodsmentioning

confidence: 99%

Determinants in the HTLV-1 capsid major homology region that are critical for virus particle assembly

Yang,

Arndt,

Zhang

et al. 2024

Preprint

Self Cite

View full text Add to dashboard Cite

The Gag protein of retroviruses is the primary driver of virus particle assembly. Particle morphologies among retroviral genera are distinct, with intriguing differences observed relative to HIV-1, particularly that of human T-cell leukemia virus type 1 (HTLV-1). In contrast to HIV-1 and other retroviruses where the capsid (CA) carboxy-terminal domain (CTD) possesses the key amino acid determinants involved in driving Gag-Gag interactions, we have previously demonstrated that the amino-terminal domain (NTD) encodes the key residues crucial for Gag multimerization and immature particle production. Here in this study, we sought to thoroughly interrogate the conserved HTLV-1 major homology region (MHR) of the CACTDto determine whether this region harbors residues important for particle assembly. In particular, site-directed mutagenesis of the HTLV-1 MHR was conducted, and mutants were analyzed for their ability to impact Gag subcellular distribution, particle production and morphology, as well as the CA-CA assembly kinetics. Several key residues (i.e., Q138, E142, Y144, F147 and R150), were found to significantly impact Gag multimerization and particle assembly. Taken together, these observations imply that while the HTLV-1 CANTDacts as the major region involved in CA-CA interactions, residues in the MHR can impact Gag multimerization, particle assembly and morphology, and likely play an important role in the conformation the CACTDthat is required for CA-CA interactions.

show abstract

Human Immunodeficiency Virus Type 2 Capsid Protein Mutagenesis Reveals Amino Acid Residues Important for Virus Particle Assembly

Cited by 5 publications

References 60 publications

HIV-2 Immature Particle Morphology Provides Insights into Gag Lattice Stability and Virus Maturation

HIV-2 Immature Particle Morphology Provides Insights into Gag Lattice Stability and Virus Maturation

HIV-2 Immature Particle Morphology Provides Insights into Gag Lattice Stability and Virus Maturation

Determinants in the HTLV-1 capsid major homology region that are critical for virus particle assembly

Contact Info

Product

Resources

About