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Nek1 regulates cell death and mitochondrial membrane permeability through phosphorylation of VDAC1

Yumay Chen1, William J Craigen, Daniel J Riley

  • 1Department of Medicine, Division of Nephrology, The University of Texas Health Science Center at San Antonio, San Antonio, Texas 78229-3900, USA. cheny@uthscsa.edu

Cell Cycle (Georgetown, Tex.)
|January 23, 2009
PubMed

Insights

NIMA-related protein kinase 1 (Nek1) prevents cell death after DNA damage by phosphorylating voltage-dependent anion channel 1 (VDAC1). Loss of Nek1 function leads to mitochondrial cell death pathway activation.

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biochemistry

Background:

  • NIMA-related protein kinase 1 (Nek1) is crucial for cellular survival following DNA damage.
  • The precise mechanism by which cells lacking functional Nek1 undergo death remains unclear.
  • Understanding Nek1's role is vital for comprehending DNA damage response pathways.

Purpose of the Study:

  • To elucidate the mechanism by which Nek1 regulates cell death after DNA damage.
  • To identify the specific molecular targets and pathways involved in Nek1-mediated cell survival.
  • To investigate the role of Nek1 in regulating mitochondrial cell death.

Main Methods:

  • Yeast two-hybrid system, GST pull-down assays, and reciprocal immunoprecipitation to study Nek1-VDAC1 interaction.
  • Mitochondrial localization studies of Nek1.
  • Expression of wild-type, kinase-dead Nek1 mutants, and VDAC1 mutants (S193A, S193E) in cells.
  • Assessment of cell viability, mitochondrial membrane permeability (MMP), and VDAC1 phosphorylation at Serine 193 (S193) after DNA damage (UV irradiation) or gene silencing.

Main Results:

  • Nek1 directly interacts with voltage-dependent anion channel 1 (VDAC1) and localizes to mitochondria.
  • Loss of Nek1 function or expression leads to decreased VDAC1-S193 phosphorylation, increased MMP, and accelerated cell death.
  • Expression of a non-phosphorylatable VDAC1-S193A mutant mimics Nek1 deficiency, causing cell death.
  • Constitutively phosphorylated VDAC1 mutant (S193E) partially rescues cell death but only transiently.

Conclusions:

  • Nek1 regulates the mitochondrial cell death pathway by phosphorylating VDAC1 at Serine 193.
  • This Nek1-VDAC1 interaction and subsequent phosphorylation is a key mechanism preventing excessive cell death after DNA damage.
  • This study provides the first direct evidence of a specific kinase regulating VDAC1 activity in the context of DNA damage response.

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