Prolyl hydroxylase 3 (PHD3) modulates catabolic effects of tumor necrosis factor-α (TNF-α) on cells of the nucleus pulposus through co-activation of nuclear factor κB (NF-κB)/p65 signaling

Nobuyuki Fujita, Shilpa S. Gogate, Kazuhiro Chiba, Yoshiaki Toyama, Irving M. Shapiro, Makarand V. Risbud

Research output: Contribution to journalArticle

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Abstract

Recent studies suggest a differential role of prolyl hydroxylase (PHD) isoforms in controlling hypoxia-inducible factor (HIF)-α degradation and activity in nucleus pulposus (NP) cells. However, the regulation and function of PHDs under inflammatory conditions that characterize disc disease are not yet known. Here, we show that in NP cells, TNF-α and IL-1β induce PHD3 expression through NF-κB. Lentiviral delivery of Sh-p65 and Sh-IKKβ confirms that cytokine-mediated PHD3 expression is NF-κB-dependent. It is noteworthy that although both cytokines induce HIF activity, mechanistic studies using Sh-HIF-1α and PHD3 promoter/enhancer constructs harboring well characterized hypoxia response element (HRE) show lack of HIF involvement in cytokine-mediated PHD3 expression. Loss-of-function studies clearly indicate that PHD3 serves as a co-activator of NF-κB signaling activity in NP cells; PHD3 interacts with, and co-localizes with, p65. We observed that when PHD3 is silenced, there is a significant decrease in TNF-α-induced expression of catabolic markers that include ADAMTS5, syndecan4, MMP13, and COX2, and at the same time, there is restoration of aggrecan and collagen type II expression. It is noteworthy that hydroxylase function of PHDs is not required for mediating cytokine-dependent gene expression. These findings show that by enhancing the activity of inflammatory cytokines, PHD3 may serve a critical role in degenerative disc disease.

Original languageEnglish
Pages (from-to)39942-39953
Number of pages12
JournalJournal of Biological Chemistry
Volume287
Issue number47
DOIs
Publication statusPublished - 2012 Nov 16

Fingerprint

Cell Nucleus
Tumor Necrosis Factor-alpha
Chemical activation
Cytokines
Hypoxia-Inducible Factor-Proline Dioxygenases
Syndecan-4
Prolyl Hydroxylases
Aggrecans
Hypoxia-Inducible Factor 1
Collagen Type II
Response Elements
Mixed Function Oxygenases
2-oxoglutarate 3-dioxygenase proline
Nucleus Pulposus
Interleukin-1
Gene expression
Restoration
Protein Isoforms
Gene Expression
Degradation

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Prolyl hydroxylase 3 (PHD3) modulates catabolic effects of tumor necrosis factor-α (TNF-α) on cells of the nucleus pulposus through co-activation of nuclear factor κB (NF-κB)/p65 signaling. / Fujita, Nobuyuki; Gogate, Shilpa S.; Chiba, Kazuhiro; Toyama, Yoshiaki; Shapiro, Irving M.; Risbud, Makarand V.

In: Journal of Biological Chemistry, Vol. 287, No. 47, 16.11.2012, p. 39942-39953.

Research output: Contribution to journalArticle

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