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Signal integration and gene induction by a functionally distinct STAT3 phosphoform.

Matthew S Waitkus1, Unni M Chandrasekharan, Belinda Willard

  • 1Department of Cellular and Molecular Medicine, Lerner Research Institute, Cleveland Clinic, Cleveland, Ohio, USA.

Molecular and Cellular Biology
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A novel STAT3 protein modification, independent of Tyr(705) phosphorylation, was discovered. This doubly phosphorylated STAT3 (pSTAT3) is regulated by GSK-3 and shows elevated levels in kidney cancer, suggesting a new therapeutic target.

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Area of Science:

  • Molecular Biology
  • Cancer Research
  • Signal Transduction

Background:

  • Aberrant Signal Transducer and Activator of Transcription 3 (STAT3) activation drives solid tumor progression and angiogenesis.
  • Canonical STAT3 function is typically assessed by Tyr(705) phosphorylation (pSTAT3).

Purpose of the Study:

  • To identify and characterize noncanonical mechanisms of STAT3 activation.
  • To investigate the role of novel STAT3 phosphoforms in cancer signaling.

Main Methods:

  • Quantitative liquid chromatography-tandem mass spectrometry (LC-MS/MS) for phosphoform discovery.
  • Functional assays involving epidermal growth factor receptor (EGFR) and protease-activated receptor 1 (PAR-1) activation in endothelial cells.
  • Gene depletion of glycogen synthase kinase 3 alpha and beta (GSK-3α/β) to assess pathway inhibition.

Main Results:

  • Discovery of a novel STAT3 phosphoform phosphorylated at Thr(714) and Ser(727) by GSK-3α/β.
  • This doubly phosphorylated STAT3 (pSTAT3) is crucial for STAT3-dependent gene induction in response to combined EGFR and PAR-1 signaling.
  • Elevated levels of doubly phosphorylated STAT3, but not Tyr(705)-phosphorylated STAT3, were observed in clear-cell renal-cell carcinoma.

Conclusions:

  • A functionally distinct, noncanonical STAT3 phosphoform regulates target gene expression in a combinatorial signaling context.
  • GSK-3α/β-STAT3 signaling represents a potential therapeutic target for renal-cell carcinoma.