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RNF219 interacts with CCR4-NOT in regulating stem cell differentiation

Hao Du1, Chen Chen1, Yan Wang1

  • 1School of Life Science and Technology, The Key Laboratory of Developmental Genes and Human Disease, Southeast University, Nanjing 210096, China.

Journal of Molecular Cell Biology
|October 26, 2020
PubMed

Insights

RING finger protein 219 (RNF219) interacts with the CCR4-NOT deadenylase complex, regulating RNA stability and gene expression. RNF219 is crucial for maintaining embryonic stem cell pluripotency and neuronal lineage commitment.

Area of Science:

  • Molecular Biology
  • Gene Regulation
  • Stem Cell Biology

Background:

  • RNA stability is critical for controlling gene expression.
  • Deadenylation is a key rate-limiting step in RNA decay.
  • The CCR4-NOT complex is a major deadenylase machinery.

Purpose of the Study:

  • To investigate the role of RING finger protein 219 (RNF219) in RNA regulation.
  • To determine RNF219's interaction with the CCR4-NOT complex.
  • To elucidate RNF219's function in embryonic stem cell (ES cell) differentiation and pluripotency.

Main Methods:

  • Co-immunoprecipitation to identify protein interactions.
  • In vitro biochemical assays to assess deadenylation activity.
  • RNA sequencing (RNA-seq) to analyze gene expression changes.
  • CRISPR/Cas9-mediated gene knockdown in mouse ES cells.

Main Results:

  • RNF219 directly interacts with the CCR4-NOT deadenylase complex.
  • The RNF219-CCR4-NOT complex demonstrates in vitro deadenylation activity.
  • RNF219 knockdown downregulates 2-cell-specific and neuronal genes in ES cells.
  • Depletion of CNOT10, a CCR4-NOT subunit, upregulates these genes.
  • RNF219 depletion impairs neuronal lineage commitment during ES cell differentiation.

Conclusions:

  • RNF219 is a novel interacting partner of the CCR4-NOT complex.
  • RNF219 plays a significant role in regulating RNA stability and gene expression.
  • RNF219 is essential for maintaining mouse ES cell pluripotency and proper differentiation.
Keywords:
CCR4–NOTRNF219deadenylationprotein complexstem cell differentiation

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