Cytosolic DNA Promotes Signal Transducer and Activator of Transcription 3 (STAT3) Phosphorylation by TANK-binding Kinase 1 (TBK1) to Restrain STAT3 Activity

J Biol Chem. 2017 Mar 31;292(13):5405-5417. doi: 10.1074/jbc.M116.771964. Epub 2017 Feb 10.

Abstract

Cytosolic DNA can elicit beneficial as well as undesirable immune responses. For example, viral or microbial DNA triggers cell-intrinsic immune responses to defend against infections, whereas aberrant cytosolic accumulation of self-DNA results in pathological conditions, such as autoimmunity. Given the importance of these DNA-provoked responses, a better understanding of their molecular mechanisms is needed. Cytosolic DNA engages stimulator of interferon genes (STING) to activate TANK-binding kinase 1 (TBK1), which subsequently phosphorylates the transcription factor interferon regulatory factor 3 (IRF3) to promote interferon expression. Recent studies have reported that additional transcription factors, including nuclear factor κB (NF-κB) and signal transducer and activator of transcription 6 (STAT6), are also activated by cytosolic DNA, suggesting that cytosolic DNA-induced gene expression is orchestrated by multiple factors. Here we show that cytosolic DNA activates STAT3, another member of the STAT family, via an autocrine mechanism involving interferon β (IFNβ) and IL-6. Additionally, we observed a novel cytosolic DNA-induced phosphorylation at serine 754 in the transactivation domain of STAT3. Upon cytosolic DNA stimulation, Ser754 is directly phosphorylated by TBK1 in a STING-dependent manner. Moreover, Ser754 phosphorylation inhibits cytosolic DNA-induced STAT3 transcriptional activity and selectively reduces STAT3 target genes that are up-regulated in response to cytosolic DNA. Taken together, our results suggest that cytosolic DNA-induced STAT3 activation via IFNβ and IL-6 is restrained by Ser754 phosphorylation of STAT3. Our findings reveal a new signaling axis downstream of the cytosolic DNA pathway and suggest potential interactions between innate immune responses and STAT3-driven oncogenic pathways.

Keywords: Cytosolic DNA; STAT transcription factor; STAT3; STING; TBK1; cell signaling; innate immunity; phosphorylation.

MeSH terms

  • Autocrine Communication
  • Cell Line
  • DNA / immunology*
  • DNA / physiology
  • Gene Expression Regulation / immunology
  • Humans
  • Immunity, Innate
  • Interferon-beta
  • Interleukin-6
  • Membrane Proteins / metabolism
  • Phosphorylation
  • Protein Serine-Threonine Kinases / metabolism*
  • STAT3 Transcription Factor / metabolism*
  • Signal Transduction*

Substances

  • Interleukin-6
  • Membrane Proteins
  • STAT3 Transcription Factor
  • STAT3 protein, human
  • STING1 protein, human
  • Interferon-beta
  • DNA
  • Protein Serine-Threonine Kinases
  • TBK1 protein, human