Signal Peptide of HIV-1 Envelope Modulates Glycosylation Impacting Exposure of V1V2 Epitopes

Upadhyay, Chitra; Feyznezhad, Roya; Cao, Liwei; Chan, Kuei-Chuan; Liu, Kevin; Yang, Weiming; Zhang, Hui; Yolitz, Jason; Arthos, James; Nádas, Arthur; Kong, Xiang‐Peng; Zolla‐Pazner, Susan; Hioe, Catarina E.

doi:10.1101/2020.07.21.212183

Cited by 6 publications

(2 citation statements)

References 99 publications

(123 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…As expected, NFL eliminated gp120 and gp41 bands (missing red and green dots, respectively in lanes 2-10). Consistent with previous studies [30,34,39,[43][44][45][46]63,64], we detected two forms of gp160: a "mature" gp160 (gp160m, magenta dots) bearing complex glycans and an "immature" gp160 (gp160i, yellow dots) bearing high mannose glycans. In NFL mutants, the gp120 band of the SOS parent (Fig.…”

Section: Expression and Maturation Of Membrane Trimer Mutantssupporting

confidence: 91%

“…On the other hand, native membrane gp120/gp41 trimers, but not UNC, are exclusively bound by NAbs [30,33,34]. Incomplete gp120/gp41 processing is a consistent unwanted feature of membrane Env expressed on VLPs, pseudovirions and even infectious molecular clones expressed in 293T cells [30,[43][44][45]. Non-functional Env comes in a variety of forms.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Impact of Common Modifications on the Antigenic Profile and Glycosylation of Membrane-Expressed HIV-1 Envelope Glycoprotein

Tong

D’Addabbo

et al. 2023

Preprint

View full text Add to dashboard Cite

Recent HIV-1 vaccine development has centered on “near native” soluble envelope glycoprotein (Env) trimers. These trimers are artificially stabilized laterally (between protomers) and apically (between gp120 and gp41). These same stabilizing mutations have been leveraged for use in membrane-expressed Env mRNA vaccines, although their precise effects in this context are unclear. To address this question, we investigated the effects of Env mutations expressed on virus-like particle (VLP) in 293T cells. Uncleaved (UNC) trimers were laterally unstable upon gentle lysis from membranes. However, gp120/gp41 processing improved lateral stability. Due to inefficient gp120/gp41 processing, UNC is incorporated into VLPs. A linker between gp120 and gp41 (NFL) neither improved trimer stability nor its antigenic profile. An artificially introduced enterokinase cleavage site allowed processing post-expression, resulting in increased trimer stability. Gp41 N-helix mutations I559P and NT1-5 both imparted lateral trimer stability, but concomitantly reduced gp120/gp41 processing and/or impacted V2 apex and interface NAb binding. I559P consistently reduced recognition by HIV+ donor plasmas, further supporting antigenic differences. Mutations in the gp120 bridging sheet failed to stabilize membrane trimers in a pre-fusion conformation, reduced gp120/gp41 processing and exposed non-neutralizing epitopes. Reduced glycan maturation and increased sequon skipping were common effect of mutations. In some cases, this may be due to increased rigidity which limits access to glycan processing enzymes. In contrast, viral gp120 did not show glycan skipping. We observed a minor species of high mannose glycan only gp160 in particle preparations. This was unaffected by any mutations and instead bypasses normal folding and glycan maturation processes. Including the full gp41 cytoplasmic tail led to markedly reduced gp120/gp41 processing and increased the proportion of high mannose gp160. Remarkably, NAbs were unable to bind to full-length Env trimers. Overall, our findings suggest caution in leveraging mutations to ensure they impart valuable membrane trimer phenotypes for vaccine use.

show abstract

Section: Expression and Maturation Of Membrane Trimer Mutantssupporting

confidence: 91%

Section: Introductionmentioning

confidence: 99%

Impact of Common Modifications on the Antigenic Profile and Glycosylation of Membrane-Expressed HIV-1 Envelope Glycoprotein

Tong

D’Addabbo

et al. 2023

Preprint

View full text Add to dashboard Cite

show abstract

The Antiviral Effects of 2-Deoxy-D-glucose (2-DG), a Dual D-Glucose and D-Mannose Mimetic, against SARS-CoV-2 and Other Highly Pathogenic Viruses

et al. 2022

View full text Add to dashboard Cite

Viral infection almost invariably causes metabolic changes in the infected cell and several types of host cells that respond to the infection. Among metabolic changes, the most prominent is the upregulated glycolysis process as the main pathway of glucose utilization. Glycolysis activation is a common mechanism of cell adaptation to several viral infections, including noroviruses, rhinoviruses, influenza virus, Zika virus, cytomegalovirus, coronaviruses and others. Such metabolic changes provide potential targets for therapeutic approaches that could reduce the impact of infection. Glycolysis inhibitors, especially 2-deoxy-D-glucose (2-DG), have been intensively studied as antiviral agents. However, 2-DG’s poor pharmacokinetic properties limit its wide clinical application. Herein, we discuss the potential of 2-DG and its novel analogs as potent promising antiviral drugs with special emphasis on targeted intracellular processes.

show abstract

Dual Role of HIV-1 Envelope Signal Peptide in Immune Evasion

Upadhyay¹,

Rao²,

Feyznezhad³

2022

Viruses

View full text Add to dashboard Cite

HIV-1 Env signal peptide (SP) is an important contributor to Env functions. Env is generated from Vpu/Env encoded bicistronic mRNA such that the 5′ end of Env-N-terminus, that encodes for Env-SP overlaps with 3′ end of Vpu. Env SP displays high sequence diversity, which translates into high variability in Vpu sequence. This study aimed to understand the effect of sequence polymorphism in the Vpu-Env overlapping region (VEOR) on the functions of two vital viral proteins: Vpu and Env. We used infectious molecular clone pNL4.3-CMU06 and swapped its SP (or VEOR) with that from other HIV-1 isolates. Swapping VEOR did not affect virus production in the absence of tetherin however, presence of tetherin significantly altered the release of virus progeny. VEOR also altered Vpu’s ability to downregulate CD4 and tetherin. We next tested the effect of these swaps on Env functions. Analyzing the binding of monoclonal antibodies to membrane embedded Env revealed changes in the antigenic landscape of swapped Envs. These swaps affected the oligosaccharide composition of Env-N-glycans as shown by changes in DC-SIGN-mediated virus transmission. Our study suggests that genetic diversity in VEOR plays an important role in the differential pathogenesis and also assist in immune evasion by altering Env epitope exposure.

show abstract

Signal Peptide of HIV-1 Envelope Modulates Glycosylation Impacting Exposure of V1V2 Epitopes

Cited by 6 publications

References 99 publications

Impact of Common Modifications on the Antigenic Profile and Glycosylation of Membrane-Expressed HIV-1 Envelope Glycoprotein

Impact of Common Modifications on the Antigenic Profile and Glycosylation of Membrane-Expressed HIV-1 Envelope Glycoprotein

The Antiviral Effects of 2-Deoxy-D-glucose (2-DG), a Dual D-Glucose and D-Mannose Mimetic, against SARS-CoV-2 and Other Highly Pathogenic Viruses

Dual Role of HIV-1 Envelope Signal Peptide in Immune Evasion

Contact Info

Product

Resources

About