Global protein phosphorylation dynamics during deoxynivalenol-induced ribotoxic stress response in the macrophage

Xiao Pan, Douglas A. Whitten, Ming Wu, Christina Chan, Curtis G. Wilkerson, James J. Pestka

    Research output: Contribution to journalArticle

    • 16 Citations

    Abstract

    Deoxynivalenol (DON), a trichothecene mycotoxin produced by Fusarium that commonly contaminates food, is capable of activating mononuclear phagocytes of the innate immune system via a process termed the ribotoxic stress response (RSR). To encapture global signaling events mediating RSR, we quantified the early temporal (≤ 30. min) phosphoproteome changes that occurred in RAW 264.7 murine macrophage during exposure to a toxicologically relevant concentration of DON (250. ng/mL). Large-scale phosphoproteomic analysis employing stable isotope labeling of amino acids in cell culture (SILAC) in conjunction with titanium dioxide chromatography revealed that DON significantly upregulated or downregulated phosphorylation of 188 proteins at both known and yet-to-be functionally characterized phosphosites. DON-induced RSR is extremely complex and goes far beyond its prior known capacity to inhibit translation and activate MAPKs. Transcriptional regulation was the main target during early DON-induced RSR, covering over 20% of the altered phosphoproteins as indicated by Gene Ontology annotation and including transcription factors/cofactors and epigenetic modulators. Other biological processes impacted included cell cycle, RNA processing, translation, ribosome biogenesis, monocyte differentiation and cytoskeleton organization. Some of these processes could be mediated by signaling networks involving MAPK-, NFκB-, AKT- and AMPK-linked pathways. Fuzzy c-means clustering revealed that DON-regulated phosphosites could be discretely classified with regard to the kinetics of phosphorylation/dephosphorylation. The cellular response networks identified provide a template for further exploration of the mechanisms of trichothecenemycotoxins and other ribotoxins, and ultimately, could contribute to improved mechanism-based human health risk assessment.

    Original languageEnglish (US)
    Pages (from-to)201-211
    Number of pages11
    JournalToxicology and Applied Pharmacology
    Volume268
    Issue number2
    DOIs
    StatePublished - Apr 5 2013

    Profile

    Macrophages
    Phosphorylation
    Proteins
    Paranasal Sinus Neoplasms
    Trichothecenes
    Isotope Labeling
    Molecular Sequence Annotation
    Gene Ontology
    AMP-Activated Protein Kinases
    Mycotoxins
    Phosphoproteins
    Fusarium
    Biogenesis
    Phagocytes
    Biological Processes
    Titanium
    Cytoskeleton
    Ribosomes
    Epigenomics
    Cluster Analysis

    Keywords

    • Deoxynivalenol
    • Phosphorylation
    • Quantitative proteomics
    • Ribotoxic stress response
    • Trichothecene mycotoxin

    ASJC Scopus subject areas

    • Pharmacology
    • Toxicology

    Cite this

    Global protein phosphorylation dynamics during deoxynivalenol-induced ribotoxic stress response in the macrophage. / Pan, Xiao; Whitten, Douglas A.; Wu, Ming; Chan, Christina; Wilkerson, Curtis G.; Pestka, James J.

    In: Toxicology and Applied Pharmacology, Vol. 268, No. 2, 05.04.2013, p. 201-211.

    Research output: Contribution to journalArticle

    Pan, Xiao; Whitten, Douglas A.; Wu, Ming; Chan, Christina; Wilkerson, Curtis G.; Pestka, James J. / Global protein phosphorylation dynamics during deoxynivalenol-induced ribotoxic stress response in the macrophage.

    In: Toxicology and Applied Pharmacology, Vol. 268, No. 2, 05.04.2013, p. 201-211.

    Research output: Contribution to journalArticle

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    abstract = "Deoxynivalenol (DON), a trichothecene mycotoxin produced by Fusarium that commonly contaminates food, is capable of activating mononuclear phagocytes of the innate immune system via a process termed the ribotoxic stress response (RSR). To encapture global signaling events mediating RSR, we quantified the early temporal (≤ 30. min) phosphoproteome changes that occurred in RAW 264.7 murine macrophage during exposure to a toxicologically relevant concentration of DON (250. ng/mL). Large-scale phosphoproteomic analysis employing stable isotope labeling of amino acids in cell culture (SILAC) in conjunction with titanium dioxide chromatography revealed that DON significantly upregulated or downregulated phosphorylation of 188 proteins at both known and yet-to-be functionally characterized phosphosites. DON-induced RSR is extremely complex and goes far beyond its prior known capacity to inhibit translation and activate MAPKs. Transcriptional regulation was the main target during early DON-induced RSR, covering over 20% of the altered phosphoproteins as indicated by Gene Ontology annotation and including transcription factors/cofactors and epigenetic modulators. Other biological processes impacted included cell cycle, RNA processing, translation, ribosome biogenesis, monocyte differentiation and cytoskeleton organization. Some of these processes could be mediated by signaling networks involving MAPK-, NFκB-, AKT- and AMPK-linked pathways. Fuzzy c-means clustering revealed that DON-regulated phosphosites could be discretely classified with regard to the kinetics of phosphorylation/dephosphorylation. The cellular response networks identified provide a template for further exploration of the mechanisms of trichothecenemycotoxins and other ribotoxins, and ultimately, could contribute to improved mechanism-based human health risk assessment.",
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