Structure-based energetics of protein interfaces guides foot-and-mouth disease virus vaccine design

Kotecha, Abhay; Seago, Julian; Scott, Katherine Anne; Burman, Alison; Loureiro, Silvia; Ren, Jingshan; Porta, Claudine; Ginn, H.; Jackson, Terry; Pérez-Martín, Eva; Siebert, C. Alistair; Paul, G.; Huiskonen, Juha T.; Jones, Ian M.; Esnouf, Robert; Fry, Elizabeth E.; Maree, Francois Frederick; Charleston, Bryan; Stuart, David I.

doi:10.1038/nsmb.3096

Cited by 94 publications

(125 citation statements)

References 54 publications

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“…Indeed these regions can be targeted to overstabilize the virus capsid, for example by capsid engineering for improved vaccines (23), or to destabilize the capsid: for example, NAb E18 binds here and causes ejection of the genome from enterovirus EV71 (13). We have found that the potent NAb R10 Fab binds in this region and somewhat destabilizes the particles, consistent with HAV uncoating also proceeding via changes at the pentamer interface.…”

Section: Discussionmentioning

confidence: 63%

Potent neutralization of hepatitis A virus reveals a receptor mimic mechanism and the receptor recognition site

Wang

Zhu

Dang

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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Hepatitis A virus (HAV) infects ∼1.4 million people annually and, although there is a vaccine, there are no licensed therapeutic drugs. HAV is unusually stable (making disinfection problematic) and little is known of how it enters cells and releases its RNA. Here we report a potent HAV-specific monoclonal antibody, R10, which neutralizes HAV infection by blocking attachment to the host cell. High-resolution cryo-EM structures of HAV full and empty particles and of the complex of HAV with R10 Fab reveal the atomic details of antibody binding and point to a receptor recognition site at the pentamer interface. These results, together with our observation that the R10 Fab destabilizes the capsid, suggest the use of a receptor mimic mechanism to neutralize virus infection, providing new opportunities for therapeutic intervention.picornavirus | entry | neutralizing mechanism | receptor recognition | uncoating H epatitis A virus (HAV) is an ancient and ubiquitous pathogen found in primates and small mammals (1). It is a picornavirus with many distinctive features: in the blood it is found in an enveloped form but is shed in feces as a naked, unenveloped particle (2); it possesses a low G/C ratio in the genome sequence and a strong codon bias (3); it grows poorly in tissue culture; it possesses a 67-residue carboxyl-terminal extension of VP1 (VP1-2A or VPX), which is important for viral assembly (4); and it has a very short, nonmyristoylated VP4 (∼23 residues) (5). Because of its unusual properties, HAV remains enigmatic and occupies an evolutionary position on the periphery of picornaviruses (6, 7).Our previous crystal structure of unenveloped HAV (7) showed no trace of the canyon that encircles the fivefold axes of enteroviruses and is often the site of receptor binding (8, 9). Indeed, the structure provided no clues as to where T-cell Ig and mucin 1 (TIM-1), the proposed receptor (10, 11), might attach. In addition, the virus capsid contains no pocket factor and can withstand remarkably high temperature and low pH, indicating an uncoating mechanism (7) unlike that of enteroviruses; heating of HAV particles does not transform virions to an expanded state in vitro. Structural characterization of a HAV-TIM-1 complex might elucidate the cell-entry mechanism, but difficulty in obtaining homogenous complex preparations because of the low binding affinity of TIM-1, at least for unenveloped particles, has made this challenging. However, it is known that neutralizing antibodies (NAbs) protect against virus infection by mechanisms that include blocking attachment to the cellular receptor, overstabilizing the virus, preventing viral genome release, or physically destabilizing the virus (12, 13). Structural studies of virus complexes with such antibodies can therefore help illuminate these underlying biological functions. Here we identify a highly potent Nab, R10, which can neutralize HAV infection efficiently by blocking attachment to the host cell. The complex structure of the HAV full particle and R10 Fab suggest where the re...

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Section: Discussionmentioning

confidence: 63%

Potent neutralization of hepatitis A virus reveals a receptor mimic mechanism and the receptor recognition site

Wang

Zhu

Dang

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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“…We recently showed that using reverse genetic approaches, we could improve stability by introducing stabilising mutations into a SAT2 infectious clone and recovering stabilised viruses [20]. Residue substitutions were predicted by comparing crystallography structures, sequence data, in silico calculations and modelling of the inter-pentameric interfaces between the thermostable A serotype and the more unstable O and SAT2 serotype viruses [20].…”

Section: Introductionmentioning

confidence: 99%

“…Recent the manipulation of the biological properties of field or laboratory strains [17][18][19] and structural analyses of the capsid have allowed for the design of thermostable mutations [20,21] integrated into reverse genetic approaches [22,23]. Several studies have shown that chimeric vaccines successfully induce protective immune responses and protect FMD host species against live virus challenge [23][24][25].…”

Section: Introductionmentioning

confidence: 99%

Evaluation of immune responses of stabilised SAT2 antigens of foot-and-mouth disease in cattle

Scott

Rathogwa

Capozzo³

et al. 2017

Vaccine

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“…Deciphering the molecular basis that define capsid stability is important not only for the understanding of acid-induced uncoating but also for the improvement of aphthovirus vaccines based on empty capsids or inactivated virions (14)(15)(16)(17)(18). Along this line, the analysis of picornavirus mutants with altered acid sensitivities has provided valuable information to understand the mechanism of acid-dependent uncoating and to understand the molecular basis of capsid stability (13,(19)(20)(21)(22)(23)(24)(25).…”

Section: Importancementioning

confidence: 98%

Equine Rhinitis A Virus Mutants with Altered Acid Resistance Unveil a Key Role of VP3 and Intrasubunit Interactions in the Control of the pH Stability of the Aphthovirus Capsid

Caridi

Cañas‐Arranz

Vázquez-Calvo

et al. 2016

J Virol

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Equine rhinitis A virus (ERAV) is a picornavirus associated with respiratory disease in horses and is genetically closely related to foot-and-mouth disease virus (FMDV), the prototype aphthovirus. ERAV has recently gained interest as an FMDV alternative for the study of aphthovirus biology, including cell entry and uncoating or antiviral testing. As described for FMDV, current data support that acidic pH inside cellular endosomes triggers ERAV uncoating. In order to provide further insights into aphthovirus uncoating mechanism, we have isolated a panel of ERAV mutants with altered acid sensitivity and that differed on their degree of sensitivity to the inhibition of endosome acidification. These results provide functional evidence of the involvement of acidic pH on ERAV uncoating within endosomes. Remarkably, all amino acid substitutions found in acid-labile or acid-resistant ERAVs were located in the capsid protein VP3, indicating that this protein plays a pivotal role for the control of pH stability of the ERAV capsid. Moreover, all amino acid substitutions mapped at the intraprotomer interface between VP3 and VP2 or between VP3 and the N terminus of VP1. These results expand our knowledge on the regions that regulate the acid stability of aphthovirus capsid and should be taken into account when using ERAV as a surrogate of FMDV. IMPORTANCEThe viral capsid constitutes a sort of dynamic nanomachine that protects the viral genome against environmental assaults while accomplishing important functions such as receptor attachment for viral entry or genome release. We have explored the molecular determinants of aphthovirus capsid stability by isolating and characterizing a panel of equine rhinitis A virus mutants that differed on their acid sensitivity. All the mutations were located within a specific region of the capsid, the intraprotomer interface among capsid proteins, thus providing new insights into the regions that control the acid stability of aphthovirus capsid. These findings could positively contribute to the development of antiviral approaches targeting aphthovirus uncoating or the refinement of vaccine strategies based on capsid stabilization. E quine rhinitis A virus (ERAV) is a picornavirus associated with febrile respiratory disease in horses (1, 2). Due to its physical properties and its identification as a respiratory pathogen, ERAV was historically classified into the genus Rhinovirus (former Equine rhinovirus-1) of the family Picornaviridae (1). However, because of its resemblance at the nucleotide sequence level and at the genomic structure to foot-and-mouth disease virus (FMDV), ERAV has been reclassified into the Aphthovirus genus (3). Consistently, ERAV is now considered the phylogenetically most closely related virus to FMDV, which actually constitutes the prototype aphthovirus (http://www.ictvonline.org/virusTaxonomy .asp). Since FMDV is an extremely contagious and pathogenic aphthovirus that can only be manipulated in high-containment biosafety level 3 facilities, the utilization o...

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Structure-based energetics of protein interfaces guides foot-and-mouth disease virus vaccine design

Cited by 94 publications

References 54 publications

Potent neutralization of hepatitis A virus reveals a receptor mimic mechanism and the receptor recognition site

Potent neutralization of hepatitis A virus reveals a receptor mimic mechanism and the receptor recognition site

Evaluation of immune responses of stabilised SAT2 antigens of foot-and-mouth disease in cattle

Equine Rhinitis A Virus Mutants with Altered Acid Resistance Unveil a Key Role of VP3 and Intrasubunit Interactions in the Control of the pH Stability of the Aphthovirus Capsid

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