The Thr205 phosphorylation site within respiratory syncytial virus matrix (M) protein modulates M oligomerization and virus production

Monika Bajorek, Leon Caly, Kim Tran, Goedele Maertens, Ralph Tripp, Eran Bacharach, Michael Teng, Reena GHILDYAL, David Jans

    Research output: Contribution to journalArticle

    17 Citations (Scopus)

    Abstract

    Human respiratory syncytial virus (RSV) is the most common cause of bronchiolitis and pneumonia in infants and the elderly worldwide; however, there is no licensed RSV vaccine or effective drug treatment available. The RSV matrix (M) protein plays key roles in virus assembly and budding, but the protein interactions that govern budding of infectious virus are not known. In this study, we focus onMprotein and identify a key phosphorylation site (Thr205) inMthat is critical for RSV infectious virus production. Recombinant virus with a nonphosphorylatable alanine (Ala) residue at the site was markedly attenuated, whereas virus with a phosphomimetic aspartate (Asp) resulted in a nonviable virus which could only be recovered with an additional mutation inM(serine to asparagine at position 220), strongly implying that Thr205 is critical for viral infectivity. Experiments in vitro showed that mutation of Thr205 does not affectMstability or the ability to form dimers but implicate an effect on higherorder oligomer assembly. In transfected and infected cells, Asp substitution of Thr205 appeared to impairMoligomerization; typical filamentous structures still formed at the plasma membrane, butMassembly during the ensuing elongation process seemed to be impaired, resulting in shorter and more branched filaments as observed using electron microscopy (EM). Our data thus imply for the first time thatMoligomerization, regulated by a negative charge at Thr205, may be critical to production of infectious RSV.

    Original languageEnglish
    Pages (from-to)6380-6393
    Number of pages14
    JournalJournal of Virology
    Volume88
    Issue number11
    DOIs
    Publication statusPublished - 2014

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    Respiratory Syncytial Viruses
    phosphorylation
    Phosphorylation
    Viruses
    Virus Release
    viruses
    Aspartic Acid
    Proteins
    Respiratory Syncytial Virus Vaccines
    proteins
    Human respiratory syncytial virus
    Virus Assembly
    Mutation
    Bronchiolitis
    Asparagine
    Alanine
    aspartic acid
    Serine
    Pneumonia
    Electron Microscopy

    Cite this

    Bajorek, Monika ; Caly, Leon ; Tran, Kim ; Maertens, Goedele ; Tripp, Ralph ; Bacharach, Eran ; Teng, Michael ; GHILDYAL, Reena ; Jans, David. / The Thr205 phosphorylation site within respiratory syncytial virus matrix (M) protein modulates M oligomerization and virus production. In: Journal of Virology. 2014 ; Vol. 88, No. 11. pp. 6380-6393.
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    title = "The Thr205 phosphorylation site within respiratory syncytial virus matrix (M) protein modulates M oligomerization and virus production",
    abstract = "Human respiratory syncytial virus (RSV) is the most common cause of bronchiolitis and pneumonia in infants and the elderly worldwide; however, there is no licensed RSV vaccine or effective drug treatment available. The RSV matrix (M) protein plays key roles in virus assembly and budding, but the protein interactions that govern budding of infectious virus are not known. In this study, we focus onMprotein and identify a key phosphorylation site (Thr205) inMthat is critical for RSV infectious virus production. Recombinant virus with a nonphosphorylatable alanine (Ala) residue at the site was markedly attenuated, whereas virus with a phosphomimetic aspartate (Asp) resulted in a nonviable virus which could only be recovered with an additional mutation inM(serine to asparagine at position 220), strongly implying that Thr205 is critical for viral infectivity. Experiments in vitro showed that mutation of Thr205 does not affectMstability or the ability to form dimers but implicate an effect on higherorder oligomer assembly. In transfected and infected cells, Asp substitution of Thr205 appeared to impairMoligomerization; typical filamentous structures still formed at the plasma membrane, butMassembly during the ensuing elongation process seemed to be impaired, resulting in shorter and more branched filaments as observed using electron microscopy (EM). Our data thus imply for the first time thatMoligomerization, regulated by a negative charge at Thr205, may be critical to production of infectious RSV.",
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    author = "Monika Bajorek and Leon Caly and Kim Tran and Goedele Maertens and Ralph Tripp and Eran Bacharach and Michael Teng and Reena GHILDYAL and David Jans",
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    The Thr205 phosphorylation site within respiratory syncytial virus matrix (M) protein modulates M oligomerization and virus production. / Bajorek, Monika; Caly, Leon; Tran, Kim; Maertens, Goedele; Tripp, Ralph; Bacharach, Eran; Teng, Michael; GHILDYAL, Reena; Jans, David.

    In: Journal of Virology, Vol. 88, No. 11, 2014, p. 6380-6393.

    Research output: Contribution to journalArticle

    TY - JOUR

    T1 - The Thr205 phosphorylation site within respiratory syncytial virus matrix (M) protein modulates M oligomerization and virus production

    AU - Bajorek, Monika

    AU - Caly, Leon

    AU - Tran, Kim

    AU - Maertens, Goedele

    AU - Tripp, Ralph

    AU - Bacharach, Eran

    AU - Teng, Michael

    AU - GHILDYAL, Reena

    AU - Jans, David

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    N2 - Human respiratory syncytial virus (RSV) is the most common cause of bronchiolitis and pneumonia in infants and the elderly worldwide; however, there is no licensed RSV vaccine or effective drug treatment available. The RSV matrix (M) protein plays key roles in virus assembly and budding, but the protein interactions that govern budding of infectious virus are not known. In this study, we focus onMprotein and identify a key phosphorylation site (Thr205) inMthat is critical for RSV infectious virus production. Recombinant virus with a nonphosphorylatable alanine (Ala) residue at the site was markedly attenuated, whereas virus with a phosphomimetic aspartate (Asp) resulted in a nonviable virus which could only be recovered with an additional mutation inM(serine to asparagine at position 220), strongly implying that Thr205 is critical for viral infectivity. Experiments in vitro showed that mutation of Thr205 does not affectMstability or the ability to form dimers but implicate an effect on higherorder oligomer assembly. In transfected and infected cells, Asp substitution of Thr205 appeared to impairMoligomerization; typical filamentous structures still formed at the plasma membrane, butMassembly during the ensuing elongation process seemed to be impaired, resulting in shorter and more branched filaments as observed using electron microscopy (EM). Our data thus imply for the first time thatMoligomerization, regulated by a negative charge at Thr205, may be critical to production of infectious RSV.

    AB - Human respiratory syncytial virus (RSV) is the most common cause of bronchiolitis and pneumonia in infants and the elderly worldwide; however, there is no licensed RSV vaccine or effective drug treatment available. The RSV matrix (M) protein plays key roles in virus assembly and budding, but the protein interactions that govern budding of infectious virus are not known. In this study, we focus onMprotein and identify a key phosphorylation site (Thr205) inMthat is critical for RSV infectious virus production. Recombinant virus with a nonphosphorylatable alanine (Ala) residue at the site was markedly attenuated, whereas virus with a phosphomimetic aspartate (Asp) resulted in a nonviable virus which could only be recovered with an additional mutation inM(serine to asparagine at position 220), strongly implying that Thr205 is critical for viral infectivity. Experiments in vitro showed that mutation of Thr205 does not affectMstability or the ability to form dimers but implicate an effect on higherorder oligomer assembly. In transfected and infected cells, Asp substitution of Thr205 appeared to impairMoligomerization; typical filamentous structures still formed at the plasma membrane, butMassembly during the ensuing elongation process seemed to be impaired, resulting in shorter and more branched filaments as observed using electron microscopy (EM). Our data thus imply for the first time thatMoligomerization, regulated by a negative charge at Thr205, may be critical to production of infectious RSV.

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    KW - Humans

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    KW - Cercopithecus aethiops

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    KW - DNA Primers/genetics

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    KW - Respiratory Syncytial Viruses/genetics

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