Conformational Change in Herpes Simplex Virus Entry Glycoproteins Detected by Dot Blot

Sari, Tri Komala; Gianopulos, Katrina A.; Nicola, Anthony V.

doi:10.1007/978-1-4939-9814-2_18

Cited by 8 publications

(5 citation statements)

References 18 publications

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“…Serial dilutions of cell-free HSV virions were prepared in serum-free DMEM with 0.2% bovine serum albumin (BSA). Samples were blotted onto a nitrocellulose membrane using a Minifold dot blot system (Whatman) (36). Five percent milk in 0.2% PBS-Tween 20 blocking buffer was added, and the membrane was gently rocked for 20 min.…”

Section: Methodsmentioning

confidence: 99%

Glycoprotein C of Herpes Simplex Virus 1 Shields Glycoprotein B from Antibody Neutralization

Sari

Gianopulos

Nicola

2020

J Virol

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Viruses have evolved strategies to avoid neutralization by the host antibody response. Herpes simplex virus (HSV) glycoprotein C (gC) functions in viral entry and binds to complement component C3b, inhibiting complement-mediated immunity. We investigated whether gC protects HSV from antibody neutralization. HSV-1 that lacks gC was more sensitive to complement-independent neutralization by a panel of gB monoclonal antibodies than a wild-type gC rescuant virus. The presence of gC decreased neutralization by 2- to 16-fold. The gB in the native envelope of HSV-1 had reduced reactivity with antibodies in comparison to gB from the gC-null virus, suggesting that gC hampers the recognition of gB epitopes in the viral particle. The protein composition of the gC-null virus, including the surface glycoproteins essential for entry, was equivalent to that of the wild type, suggesting that gC is directly responsible for the reduced antibody recognition and neutralization. The neutralizing activity of antibodies to gD and gH antibodies was also increased in HSV lacking gC. Together, the data suggest that HSV-1 gC protects the viral envelope glycoproteins essential for entry, including gB, by shielding them from neutralization as a potential mechanism of immune evasion. IMPORTANCE HSV-1 causes lifelong infection in the human population and can be fatal in neonatal and immunocompromised individuals. There is no vaccine or cure, in part due to the ability of HSV to escape the host immune response by various mechanisms. The HSV particle contains at least 15 envelope proteins, four of which are required for entry and replication. This work suggests a novel role for gC in shielding the HSV entry glycoproteins. gC may function to help HSV escape neutralization by antibodies.

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Section: Methodsmentioning

confidence: 99%

Glycoprotein C of Herpes Simplex Virus 1 Shields Glycoprotein B from Antibody Neutralization

Sari

Gianopulos

Nicola

2020

J Virol

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“…Virions were adjusted with HCl to achieve pHs ranging from 5.0 to 7.3. Approximately 10 7 genome copies of HSV-1 were incubated at 37°C for 10 min and were then either blotted directly to a nitrocellulose membrane using a Minifold dot blot system (Whatman) or were first neutralized by addition of pretitrated amounts of 0.05 N NaOH (80). Virus-dotted nitrocellulose membranes were blocked and then incubated with antibodies to gB at neutral pH.…”

Section: Cellsmentioning

confidence: 99%

Herpes Simplex Virus Glycoprotein C Regulates Low-pH Entry

Sari

Gianopulos

Weed

et al. 2020

mSphere

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Herpes simplex viruses (HSVs) cause significant morbidity and mortality in humans worldwide. Herpesviruses mediate entry by a multicomponent virus-encoded machinery. Herpesviruses enter cells by endosomal low-pH and pH-neutral mechanisms in a cell-specific manner. HSV mediates cell entry via the envelope glycoproteins gB and gD and the heterodimer gH/gL regardless of pH or endocytosis requirements. Specifics concerning HSV envelope proteins that function selectively in a given entry pathway have been elusive. Here, we demonstrate that gC regulates cell entry and infection by a low-pH pathway. Conformational changes in the core herpesviral fusogen gB are critical for membrane fusion. The presence of gC conferred a higher pH threshold for acid-induced antigenic changes in gB. Thus, gC may selectively facilitate low-pH entry by regulating conformational changes in the fusion protein gB. We propose that gC modulates the HSV fusion machinery during entry into pathophysiologically relevant cells, such as human epidermal keratinocytes. IMPORTANCE Herpesviruses are ubiquitous pathogens that cause lifelong latent infections and that are characterized by multiple entry pathways. We propose that herpes simplex virus (HSV) gC plays a selective role in modulating HSV entry, such as entry into epithelial cells, by a low-pH pathway. gC facilitates a conformational change of the main fusogen gB, a class III fusion protein. We propose a model whereby gC functions with gB, gD, and gH/gL to allow low-pH entry. In the absence of gC, HSV entry occurs at a lower pH, coincident with trafficking to a lower pH compartment where gB changes occur at more acidic pHs. This report identifies a new function for gC and provides novel insight into the complex mechanism of HSV entry and fusion.

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“…gC contributes to entry by binding to cell surface glycosaminoglycans ( 48 ). Recent work has shown that gC can regulate entry via a low-pH pathway and impact gB conformational changes ( 49 , 50 ). Whether these gC and gH mutations contributed to the enhanced entry observed in these passaged viruses may be the subject of future work.…”

Section: Discussionmentioning

confidence: 99%

Multiple Sites on Glycoprotein H (gH) Functionally Interact with the gB Fusion Protein to Promote Fusion during Herpes Simplex Virus (HSV) Entry

Fan

Hippler

Yang

et al. 2023

mBio

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Enveloped viruses enter cells by fusing their envelope with the host cell membrane. Herpes simplex virus 1 (HSV-1) entry requires the coordinated interaction of several viral glycoproteins, including gH/gL and gB. gH/gL and gB are essential for virus replication and both proteins are targets of neutralizing antibodies. gB fuses the membranes after being activated by gH/gL, but the details of how gH/gL activates gB are not known.

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Conformational Change in Herpes Simplex Virus Entry Glycoproteins Detected by Dot Blot

Cited by 8 publications

References 18 publications

Glycoprotein C of Herpes Simplex Virus 1 Shields Glycoprotein B from Antibody Neutralization

Glycoprotein C of Herpes Simplex Virus 1 Shields Glycoprotein B from Antibody Neutralization

Herpes Simplex Virus Glycoprotein C Regulates Low-pH Entry

Multiple Sites on Glycoprotein H (gH) Functionally Interact with the gB Fusion Protein to Promote Fusion during Herpes Simplex Virus (HSV) Entry

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