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

    53 Scopus citations

    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

    ASJC Scopus subject areas

    • Biochemistry
    • Immunology
    • Hematology
    • Cell Biology

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