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Serine 347 Phosphorylation by JNKs Negatively Regulates OCT4 Protein Stability in Mouse Embryonic Stem Cells

Ki Beom Bae1, Dong Hoon Yu1, Kun Yeong Lee1

  • 1The Hormel Institute, University of Minnesota, 801 16(th) Avenue NE, Austin, MN 55912, USA.

Stem Cell Reports
|November 21, 2017
PubMed

Insights

Phosphorylation of OCT4 by JNKs at serine 347 reduces its transcriptional activity and stability, impacting embryonic stem cell self-renewal and induced pluripotent stem cell generation.

Area of Science:

  • Stem cell biology
  • Molecular and cell biology
  • Epigenetics and gene regulation

Background:

  • OCT4 is a key transcription factor essential for maintaining pluripotency in embryonic stem cells (ESCs) and reprogramming somatic cells into induced pluripotent stem cells (iPSCs).
  • The precise molecular mechanisms governing OCT4's function, particularly post-translational modifications like phosphorylation, are not fully elucidated.
  • Understanding OCT4 regulation is crucial for advancing regenerative medicine and stem cell therapies.

Purpose of the Study:

  • To investigate the role of OCT4 phosphorylation in regulating the biological functions of ESCs.
  • To identify the kinases involved in OCT4 phosphorylation and the specific phosphorylation sites.
  • To determine the impact of OCT4 phosphorylation on its transcriptional activity, protein stability, and cellular functions.

Main Methods:

  • Co-immunoprecipitation assays to detect interactions between JNKs and OCT4.
  • In vitro kinase assays to confirm direct phosphorylation of OCT4 by JNKs.
  • Western blotting and mass spectrometry to analyze OCT4 phosphorylation status and protein stability.
  • Reporter assays to measure OCT4 transcriptional activity.
  • Cell differentiation assays and iPSC generation efficiency assessments.

Main Results:

  • c-Jun N-terminal kinases (JNKs) were identified as direct interactors and phosphorylators of OCT4 at serine 347.
  • Phosphorylation of OCT4 at S347 inhibited its transcriptional activity.
  • Phosphorylated OCT4 (S347) exhibited increased binding to FBXW8, leading to reduced protein stability and enhanced proteasomal degradation.
  • A mutant OCT4 (S347A) showed delayed differentiation in mouse ESCs and enhanced iPSC generation efficiency.

Conclusions:

  • OCT4 phosphorylation at serine 347 by JNKs is a critical regulatory mechanism controlling OCT4's stability and transcriptional function.
  • This phosphorylation event plays a significant role in regulating ESC self-renewal and the efficiency of iPSC generation.
  • Targeting OCT4 phosphorylation could offer novel strategies for stem cell manipulation and therapeutic applications.
Keywords:
FBXW8JNKOCT4embryonic stem cellsinduced pluripotent stem cellsphosphorylationself-renewalubiquitination

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