Thermostability of the Foot-and-Mouth Disease Virus Capsid Is Modulated by Lethal and Viability-Restoring Compensatory Amino Acid Substitutions

Silvia López-Argüello, Verónica Rincón, Alicia Rodríguez-Huete, Encarnación Martínez-Salas, Graham Belsham, Alejandro Valbuena*, Mauricio G Mateu

*Corresponding author for this work

    Research output: Contribution to journalJournal articleResearchpeer-review

    30 Downloads (Pure)

    Abstract

    Infection by viruses depends on a balance between capsid stability and dynamics. This study has investigated biologically and biotechnologically relevant aspects of the relationship in foot-and-mouth disease virus (FMDV) between capsid structure and thermostability, and between thermostability and infectivity. In the FMDV capsid a substantial number of amino acid side chains at the interfaces between pentameric subunits are charged at neutral pH. Here a mutational analysis revealed that the essential role for virus infection of most of the 8 tested charged groups is not related to substantial changes in capsid protein expression or processing, or in capsid assembly or stability against thermally-induced dissociation into pentamers. However, the positively charged side chains of R2018 and H3141, located at the interpentameric interfaces close to the capsid 2-fold symmetry axes, were found to be critical both for virus infectivity and for keeping the capsid in a state of weak thermostability. A charge-restoring substitution, N2019H, that was repeatedly fixed during amplification of viral genomes carrying deleterious mutations, reverted both the lethal and capsid-stabilizing effects of substitution H3141A, leading to a double mutant virus with close to normal infectivity and thermolability. H3141A and other thermostabilizing substitutions had no detectable effect on capsid resistance to acid-induced dissociation into pentamers. The results suggest that FMDV infectivity requires limited local stability around the 2-fold axes at the interpentameric interfaces of the capsid. The implications for the mechanism of genome uncoating in FMDV and the development of thermostabilized vaccines against foot-and-mouth disease are discussed. IMPORTANCE This study provides novel insights into the little known structural determinants of the balance between thermal stability and instability in the capsid of foot-and-mouth disease virus, and into the relationship between capsid stability and virus infectivity. The results provide new guidelines for the development of thermostabilized empty capsid-based recombinant vaccines against foot-and-mouth disease, one of the economically most important animal diseases worldwide.
    Original languageEnglish
    Article numbere02293-18
    JournalJournal of Virology
    Volume93
    Issue number10
    Number of pages19
    ISSN0022-538X
    DOIs
    Publication statusPublished - 2019

    Keywords

    • Capsid
    • Foot-and-mouth disease virus
    • Protein engineering
    • Thermal stability
    • Vaccine

    Cite this

    López-Argüello, S., Rincón, V., Rodríguez-Huete, A., Martínez-Salas, E., Belsham, G., Valbuena, A., & Mateu, M. G. (2019). Thermostability of the Foot-and-Mouth Disease Virus Capsid Is Modulated by Lethal and Viability-Restoring Compensatory Amino Acid Substitutions. Journal of Virology, 93(10), [e02293-18]. https://doi.org/10.1128/JVI.02293-18
    López-Argüello, Silvia ; Rincón, Verónica ; Rodríguez-Huete, Alicia ; Martínez-Salas, Encarnación ; Belsham, Graham ; Valbuena, Alejandro ; Mateu, Mauricio G. / Thermostability of the Foot-and-Mouth Disease Virus Capsid Is Modulated by Lethal and Viability-Restoring Compensatory Amino Acid Substitutions. In: Journal of Virology. 2019 ; Vol. 93, No. 10.
    @article{0554c2d999474389a47a777679cea559,
    title = "Thermostability of the Foot-and-Mouth Disease Virus Capsid Is Modulated by Lethal and Viability-Restoring Compensatory Amino Acid Substitutions",
    abstract = "Infection by viruses depends on a balance between capsid stability and dynamics. This study has investigated biologically and biotechnologically relevant aspects of the relationship in foot-and-mouth disease virus (FMDV) between capsid structure and thermostability, and between thermostability and infectivity. In the FMDV capsid a substantial number of amino acid side chains at the interfaces between pentameric subunits are charged at neutral pH. Here a mutational analysis revealed that the essential role for virus infection of most of the 8 tested charged groups is not related to substantial changes in capsid protein expression or processing, or in capsid assembly or stability against thermally-induced dissociation into pentamers. However, the positively charged side chains of R2018 and H3141, located at the interpentameric interfaces close to the capsid 2-fold symmetry axes, were found to be critical both for virus infectivity and for keeping the capsid in a state of weak thermostability. A charge-restoring substitution, N2019H, that was repeatedly fixed during amplification of viral genomes carrying deleterious mutations, reverted both the lethal and capsid-stabilizing effects of substitution H3141A, leading to a double mutant virus with close to normal infectivity and thermolability. H3141A and other thermostabilizing substitutions had no detectable effect on capsid resistance to acid-induced dissociation into pentamers. The results suggest that FMDV infectivity requires limited local stability around the 2-fold axes at the interpentameric interfaces of the capsid. The implications for the mechanism of genome uncoating in FMDV and the development of thermostabilized vaccines against foot-and-mouth disease are discussed. IMPORTANCE This study provides novel insights into the little known structural determinants of the balance between thermal stability and instability in the capsid of foot-and-mouth disease virus, and into the relationship between capsid stability and virus infectivity. The results provide new guidelines for the development of thermostabilized empty capsid-based recombinant vaccines against foot-and-mouth disease, one of the economically most important animal diseases worldwide.",
    keywords = "Capsid, Foot-and-mouth disease virus, Protein engineering, Thermal stability, Vaccine",
    author = "Silvia L{\'o}pez-Arg{\"u}ello and Ver{\'o}nica Rinc{\'o}n and Alicia Rodr{\'i}guez-Huete and Encarnaci{\'o}n Mart{\'i}nez-Salas and Graham Belsham and Alejandro Valbuena and Mateu, {Mauricio G}",
    year = "2019",
    doi = "10.1128/JVI.02293-18",
    language = "English",
    volume = "93",
    journal = "Journal of Virology",
    issn = "0022-538X",
    publisher = "American Society for Microbiology",
    number = "10",

    }

    López-Argüello, S, Rincón, V, Rodríguez-Huete, A, Martínez-Salas, E, Belsham, G, Valbuena, A & Mateu, MG 2019, 'Thermostability of the Foot-and-Mouth Disease Virus Capsid Is Modulated by Lethal and Viability-Restoring Compensatory Amino Acid Substitutions', Journal of Virology, vol. 93, no. 10, e02293-18. https://doi.org/10.1128/JVI.02293-18

    Thermostability of the Foot-and-Mouth Disease Virus Capsid Is Modulated by Lethal and Viability-Restoring Compensatory Amino Acid Substitutions. / López-Argüello, Silvia; Rincón, Verónica; Rodríguez-Huete, Alicia; Martínez-Salas, Encarnación; Belsham, Graham ; Valbuena, Alejandro; Mateu, Mauricio G.

    In: Journal of Virology, Vol. 93, No. 10, e02293-18, 2019.

    Research output: Contribution to journalJournal articleResearchpeer-review

    TY - JOUR

    T1 - Thermostability of the Foot-and-Mouth Disease Virus Capsid Is Modulated by Lethal and Viability-Restoring Compensatory Amino Acid Substitutions

    AU - López-Argüello, Silvia

    AU - Rincón, Verónica

    AU - Rodríguez-Huete, Alicia

    AU - Martínez-Salas, Encarnación

    AU - Belsham, Graham

    AU - Valbuena, Alejandro

    AU - Mateu, Mauricio G

    PY - 2019

    Y1 - 2019

    N2 - Infection by viruses depends on a balance between capsid stability and dynamics. This study has investigated biologically and biotechnologically relevant aspects of the relationship in foot-and-mouth disease virus (FMDV) between capsid structure and thermostability, and between thermostability and infectivity. In the FMDV capsid a substantial number of amino acid side chains at the interfaces between pentameric subunits are charged at neutral pH. Here a mutational analysis revealed that the essential role for virus infection of most of the 8 tested charged groups is not related to substantial changes in capsid protein expression or processing, or in capsid assembly or stability against thermally-induced dissociation into pentamers. However, the positively charged side chains of R2018 and H3141, located at the interpentameric interfaces close to the capsid 2-fold symmetry axes, were found to be critical both for virus infectivity and for keeping the capsid in a state of weak thermostability. A charge-restoring substitution, N2019H, that was repeatedly fixed during amplification of viral genomes carrying deleterious mutations, reverted both the lethal and capsid-stabilizing effects of substitution H3141A, leading to a double mutant virus with close to normal infectivity and thermolability. H3141A and other thermostabilizing substitutions had no detectable effect on capsid resistance to acid-induced dissociation into pentamers. The results suggest that FMDV infectivity requires limited local stability around the 2-fold axes at the interpentameric interfaces of the capsid. The implications for the mechanism of genome uncoating in FMDV and the development of thermostabilized vaccines against foot-and-mouth disease are discussed. IMPORTANCE This study provides novel insights into the little known structural determinants of the balance between thermal stability and instability in the capsid of foot-and-mouth disease virus, and into the relationship between capsid stability and virus infectivity. The results provide new guidelines for the development of thermostabilized empty capsid-based recombinant vaccines against foot-and-mouth disease, one of the economically most important animal diseases worldwide.

    AB - Infection by viruses depends on a balance between capsid stability and dynamics. This study has investigated biologically and biotechnologically relevant aspects of the relationship in foot-and-mouth disease virus (FMDV) between capsid structure and thermostability, and between thermostability and infectivity. In the FMDV capsid a substantial number of amino acid side chains at the interfaces between pentameric subunits are charged at neutral pH. Here a mutational analysis revealed that the essential role for virus infection of most of the 8 tested charged groups is not related to substantial changes in capsid protein expression or processing, or in capsid assembly or stability against thermally-induced dissociation into pentamers. However, the positively charged side chains of R2018 and H3141, located at the interpentameric interfaces close to the capsid 2-fold symmetry axes, were found to be critical both for virus infectivity and for keeping the capsid in a state of weak thermostability. A charge-restoring substitution, N2019H, that was repeatedly fixed during amplification of viral genomes carrying deleterious mutations, reverted both the lethal and capsid-stabilizing effects of substitution H3141A, leading to a double mutant virus with close to normal infectivity and thermolability. H3141A and other thermostabilizing substitutions had no detectable effect on capsid resistance to acid-induced dissociation into pentamers. The results suggest that FMDV infectivity requires limited local stability around the 2-fold axes at the interpentameric interfaces of the capsid. The implications for the mechanism of genome uncoating in FMDV and the development of thermostabilized vaccines against foot-and-mouth disease are discussed. IMPORTANCE This study provides novel insights into the little known structural determinants of the balance between thermal stability and instability in the capsid of foot-and-mouth disease virus, and into the relationship between capsid stability and virus infectivity. The results provide new guidelines for the development of thermostabilized empty capsid-based recombinant vaccines against foot-and-mouth disease, one of the economically most important animal diseases worldwide.

    KW - Capsid

    KW - Foot-and-mouth disease virus

    KW - Protein engineering

    KW - Thermal stability

    KW - Vaccine

    U2 - 10.1128/JVI.02293-18

    DO - 10.1128/JVI.02293-18

    M3 - Journal article

    VL - 93

    JO - Journal of Virology

    JF - Journal of Virology

    SN - 0022-538X

    IS - 10

    M1 - e02293-18

    ER -