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Nuclear phosphoproteomics analysis reveals that CDK1/2 are involved in EGF-regulated constitutive pre-mRNA splicing

Xianwei Chen1, Dan Guo2, Yinghui Zhu1

  • 1CAS Key Laboratory of Genome Sciences and Information, China Gastrointestinal Cancer Research Center, Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing, 100101, China; University of Chinese Academy of Sciences, Beijing 100049, China.

Journal of Proteomics
|April 26, 2016
PubMed
Abstract

Insights

Epidermal growth factor (EGF) stimulation activates the epidermal growth factor receptor (EGFR) pathway, increasing constitutive pre-mRNA splicing. This process involves cyclin-dependent kinases 1 and 2 (CDK1/2) in MDA-MB-468 cancer cells.

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Cancer Signaling

Background:

  • The epidermal growth factor (EGF) receptor (EGFR) pathway is frequently dysregulated in cancer.
  • While EGFR's role in cytoplasmic signaling is well-studied, its nuclear functions, particularly phosphorylation responses, remain less understood.
  • EGFR signaling influences nuclear functions, impacting processes like RNA processing and cell cycle regulation.

Purpose of the Study:

  • To investigate nuclear phosphorylation changes in response to EGF stimulation.
  • To elucidate the role of EGFR in regulating pre-mRNA splicing.
  • To identify key kinases involved in EGF-mediated pre-mRNA splicing.

Main Methods:

  • Phosphoproteomics analysis of MDA-MB-468 cells following EGF stimulation.
  • Functional category analysis of identified phosphoproteins.
  • Pre-mRNA splicing assays using a β-globin minigene.
  • Inhibition and knockdown of CDK1/2 to assess their role in splicing.

Main Results:

  • EGF stimulation led to increased phosphorylation of 90 nuclear proteins involved in RNA processing and splicing.
  • EGFR activation was shown to induce constitutive pre-mRNA splicing in MDA-MB-468 cells.
  • CDK1/2 were identified as key kinases phosphorylating splicing factors, and their inhibition/knockdown reduced EGF-regulated splicing.

Conclusions:

  • EGFR activation promotes constitutive pre-mRNA splicing, extending its known regulatory roles in mRNA maturation.
  • CDK1/2 play a crucial role in mediating EGF-induced constitutive pre-mRNA splicing.
  • This study provides novel insights into the molecular mechanisms linking EGFR signaling to RNA splicing regulation.
Keywords:
CDK1/2EGFMDA-MB-468 cellsNuclear phosphoproteomicsPre-mRNA splicing

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