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Binding of 14-3-3 reader proteins to phosphorylated DNMT1 facilitates aberrant DNA methylation and gene expression

Pierre-Olivier Estève1, Guoqiang Zhang1, V K Chaithanya Ponnaluri1

  • 1New England Biolabs Inc, 240 County Road, Ipswich, MA 01938, USA.

Nucleic Acids Research
|November 11, 2015
PubMed

Insights

14-3-3 proteins bind to phosphorylated DNA methyltransferase 1 (DNMT1pSer143), inhibiting its activity. This leads to genome hypomethylation, altered gene expression, and increased cell invasion, with implications for breast cancer patient survival.

Area of Science:

  • Epigenetics
  • Molecular Biology
  • Cancer Biology

Background:

  • DNA methyltransferase 1 (DNMT1) is crucial for maintaining DNA methylation patterns during replication.
  • Phosphorylation of DNMT1 at Serine 143 (pSer143) stabilizes the enzyme.
  • 14-3-3 proteins are known to interact with phosphorylated substrates.

Purpose of the Study:

  • To investigate the role of 14-3-3 proteins as readers of DNMT1pSer143.
  • To determine the impact of 14-3-3 binding on DNMT1 activity and DNA methylation.
  • To explore the functional consequences of 14-3-3 mediated DNMT1 regulation in cellular processes and cancer.

Main Methods:

  • Co-localization and binding assays of 14-3-3 and DNMT1pSer143 in mammalian cells.
  • In vitro assays to measure DNMT1 methylation activity upon 14-3-3 binding.
  • Overexpression and depletion studies of 14-3-3 in NIH3T3 cells.
  • Analysis of gene expression and methylation patterns.
  • TCGA breast cancer patient data analysis.

Main Results:

  • 14-3-3 proteins bind to DNMT1pSer143 post-replication in mammalian cells.
  • Binding of 14-3-3 inhibits DNMT1 DNA methylation activity in vitro.
  • Overexpression of 14-3-3 leads to genome-wide hypomethylation and altered expression of genes involved in cell migration and proliferation.
  • Overexpression of 14-3-3 enhances cell invasion.
  • Increased 14-3-3 expression correlates with DNA hypomethylation and reduced survival in breast cancer patients.

Conclusions:

  • 14-3-3 acts as a reader protein for DNMT1pSer143, regulating its enzymatic activity.
  • 14-3-3-mediated inhibition of DNMT1 contributes to aberrant DNA methylation and gene expression.
  • These epigenetic alterations driven by 14-3-3 are linked to increased cell invasion and poorer prognosis in breast cancer.

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