Role of double-stranded RNA-dependent protein kinase in mediating hypersensitivity of fanconi anemia complementation group C cells to interferon γ, tumor necrosis factor-α, and double-stranded RNA

Qishen Pang, Winifred Keeble, Jane Diaz, Tracy A. Christianson, Sara Fagerlie, Keaney Rathbun, Gregory R. Faulkner, Michael O'Dwyer, Grover C. Bagby

    Research output: Contribution to journalArticle

    52 Citations (Scopus)

    Abstract

    Hematopoietic cells bearing inactivating mutations of Fanconi anemia group C (FANCC) are excessively apoptotic and demonstrate hypersensitivity not only to cross-linking agents but also to interferon γ (IFN-γ) and tumor necrosis factor-α. Seeking essential signaling pathways for this phenotype, this study quantified constitutive and induced RNA-dependent protein kinase (PKR) activation in Fanconi anemia cells of the C complementation group (FA-C). PKR was constitutively phosphorylated and exhibited an increased binding affinity for double-stranded RNA (dsRNA) in FANCC-/- cells. FANCC-/- cells were hypersensitive to both dsRNA and the combination of dsRNA and IFN-γ in that these agents induced a higher fraction of apoptosis in FANCC-/- cells than in normal cells. Overexpression of wild-type PKR-sensitized FANCC-/- cells to apoptosis induced by IFN-γ and dsRNA. Conversely, inhibition of PKR function by enforced expression of a dominant-negative inhibitory mutant of PKR (PKRΔ6) substantially reduced the IFN and dsRNA hypersensitivity of FANCC-/- cells. Two PKR target molecules, IκB-α and IRF-1, were not differentially activated in FANCC-/- cells, but enforced expression of a nonphosphorylatable form of eukaryotic translation initiation factor-2α reversed the PKR-mediated block of messenger RNA translation and partially abrogated the PKR-mediated apoptosis in FANCC-/- cells. Because no evidence was found of a PKR/FANCC complex in normal cells, it was concluded that an essential function of FANCC is to suppress, indirectly, the activity of PKR and that FANCC inactivation results in IFN hypersensitivity, at least in part, because this function of FANCC is abrogated.

    Original languageEnglish (US)
    Pages (from-to)1644-1652
    Number of pages9
    JournalBlood
    Volume97
    Issue number6
    DOIs
    StatePublished - Mar 15 2001

    Fingerprint

    eIF-2 Kinase
    Fanconi Anemia
    Double-Stranded RNA
    Interferons
    Hypersensitivity
    Tumor Necrosis Factor-alpha
    Apoptosis
    Bearings (structural)
    Prokaryotic Initiation Factor-2
    Eukaryotic Initiation Factors
    Chemical activation
    Messenger RNA
    Molecules
    Eukaryotic Initiation Factor-2

    ASJC Scopus subject areas

    • Hematology

    Cite this

    Role of double-stranded RNA-dependent protein kinase in mediating hypersensitivity of fanconi anemia complementation group C cells to interferon γ, tumor necrosis factor-α, and double-stranded RNA. / Pang, Qishen; Keeble, Winifred; Diaz, Jane; Christianson, Tracy A.; Fagerlie, Sara; Rathbun, Keaney; Faulkner, Gregory R.; O'Dwyer, Michael; Bagby, Grover C.

    In: Blood, Vol. 97, No. 6, 15.03.2001, p. 1644-1652.

    Research output: Contribution to journalArticle

    Pang, Qishen ; Keeble, Winifred ; Diaz, Jane ; Christianson, Tracy A. ; Fagerlie, Sara ; Rathbun, Keaney ; Faulkner, Gregory R. ; O'Dwyer, Michael ; Bagby, Grover C. / Role of double-stranded RNA-dependent protein kinase in mediating hypersensitivity of fanconi anemia complementation group C cells to interferon γ, tumor necrosis factor-α, and double-stranded RNA. In: Blood. 2001 ; Vol. 97, No. 6. pp. 1644-1652.
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    abstract = "Hematopoietic cells bearing inactivating mutations of Fanconi anemia group C (FANCC) are excessively apoptotic and demonstrate hypersensitivity not only to cross-linking agents but also to interferon γ (IFN-γ) and tumor necrosis factor-α. Seeking essential signaling pathways for this phenotype, this study quantified constitutive and induced RNA-dependent protein kinase (PKR) activation in Fanconi anemia cells of the C complementation group (FA-C). PKR was constitutively phosphorylated and exhibited an increased binding affinity for double-stranded RNA (dsRNA) in FANCC-/- cells. FANCC-/- cells were hypersensitive to both dsRNA and the combination of dsRNA and IFN-γ in that these agents induced a higher fraction of apoptosis in FANCC-/- cells than in normal cells. Overexpression of wild-type PKR-sensitized FANCC-/- cells to apoptosis induced by IFN-γ and dsRNA. Conversely, inhibition of PKR function by enforced expression of a dominant-negative inhibitory mutant of PKR (PKRΔ6) substantially reduced the IFN and dsRNA hypersensitivity of FANCC-/- cells. Two PKR target molecules, IκB-α and IRF-1, were not differentially activated in FANCC-/- cells, but enforced expression of a nonphosphorylatable form of eukaryotic translation initiation factor-2α reversed the PKR-mediated block of messenger RNA translation and partially abrogated the PKR-mediated apoptosis in FANCC-/- cells. Because no evidence was found of a PKR/FANCC complex in normal cells, it was concluded that an essential function of FANCC is to suppress, indirectly, the activity of PKR and that FANCC inactivation results in IFN hypersensitivity, at least in part, because this function of FANCC is abrogated.",
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    AU - Pang, Qishen

    AU - Keeble, Winifred

    AU - Diaz, Jane

    AU - Christianson, Tracy A.

    AU - Fagerlie, Sara

    AU - Rathbun, Keaney

    AU - Faulkner, Gregory R.

    AU - O'Dwyer, Michael

    AU - Bagby, Grover C.

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    N2 - Hematopoietic cells bearing inactivating mutations of Fanconi anemia group C (FANCC) are excessively apoptotic and demonstrate hypersensitivity not only to cross-linking agents but also to interferon γ (IFN-γ) and tumor necrosis factor-α. Seeking essential signaling pathways for this phenotype, this study quantified constitutive and induced RNA-dependent protein kinase (PKR) activation in Fanconi anemia cells of the C complementation group (FA-C). PKR was constitutively phosphorylated and exhibited an increased binding affinity for double-stranded RNA (dsRNA) in FANCC-/- cells. FANCC-/- cells were hypersensitive to both dsRNA and the combination of dsRNA and IFN-γ in that these agents induced a higher fraction of apoptosis in FANCC-/- cells than in normal cells. Overexpression of wild-type PKR-sensitized FANCC-/- cells to apoptosis induced by IFN-γ and dsRNA. Conversely, inhibition of PKR function by enforced expression of a dominant-negative inhibitory mutant of PKR (PKRΔ6) substantially reduced the IFN and dsRNA hypersensitivity of FANCC-/- cells. Two PKR target molecules, IκB-α and IRF-1, were not differentially activated in FANCC-/- cells, but enforced expression of a nonphosphorylatable form of eukaryotic translation initiation factor-2α reversed the PKR-mediated block of messenger RNA translation and partially abrogated the PKR-mediated apoptosis in FANCC-/- cells. Because no evidence was found of a PKR/FANCC complex in normal cells, it was concluded that an essential function of FANCC is to suppress, indirectly, the activity of PKR and that FANCC inactivation results in IFN hypersensitivity, at least in part, because this function of FANCC is abrogated.

    AB - Hematopoietic cells bearing inactivating mutations of Fanconi anemia group C (FANCC) are excessively apoptotic and demonstrate hypersensitivity not only to cross-linking agents but also to interferon γ (IFN-γ) and tumor necrosis factor-α. Seeking essential signaling pathways for this phenotype, this study quantified constitutive and induced RNA-dependent protein kinase (PKR) activation in Fanconi anemia cells of the C complementation group (FA-C). PKR was constitutively phosphorylated and exhibited an increased binding affinity for double-stranded RNA (dsRNA) in FANCC-/- cells. FANCC-/- cells were hypersensitive to both dsRNA and the combination of dsRNA and IFN-γ in that these agents induced a higher fraction of apoptosis in FANCC-/- cells than in normal cells. Overexpression of wild-type PKR-sensitized FANCC-/- cells to apoptosis induced by IFN-γ and dsRNA. Conversely, inhibition of PKR function by enforced expression of a dominant-negative inhibitory mutant of PKR (PKRΔ6) substantially reduced the IFN and dsRNA hypersensitivity of FANCC-/- cells. Two PKR target molecules, IκB-α and IRF-1, were not differentially activated in FANCC-/- cells, but enforced expression of a nonphosphorylatable form of eukaryotic translation initiation factor-2α reversed the PKR-mediated block of messenger RNA translation and partially abrogated the PKR-mediated apoptosis in FANCC-/- cells. Because no evidence was found of a PKR/FANCC complex in normal cells, it was concluded that an essential function of FANCC is to suppress, indirectly, the activity of PKR and that FANCC inactivation results in IFN hypersensitivity, at least in part, because this function of FANCC is abrogated.

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