The Anti-Apoptotic Properties of APEX1 in the Endothelium Require the First 20 Amino Acids and Converge on Thioredoxin-1

Antioxid Redox Signal. 2017 Apr 20;26(12):616-629. doi: 10.1089/ars.2016.6799. Epub 2017 Feb 22.

Abstract

The APEX nuclease (multifunctional DNA repair enzyme) 1 (APEX1) has a disordered N-terminus, a redox, and a DNA repair domain. APEX1 has anti-apoptotic properties, which have been linked to both domains depending on cell type and experimental conditions.

Aims: As protection against apoptosis is a hallmark of vessel integrity, we wanted to elucidate whether APEX1 acts anti-apoptotic in primary human endothelial cells and, if so, what the underlying mechanisms are.

Results: APEX1 inhibits apoptosis in endothelial cells by reducing Cathepsin D (CatD) cleavage, potentially by binding to the unprocessed form. Diminished CatD activation results in increased Thioredoxin-1 protein levels leading to reduced Caspase 3 activation. Consequently, apoptosis rates are decreased. This depends on the first twenty amino acids in APEX1, because APEX1 (21-318) induces CatD activity, decreases Thioredoxin-1 protein levels, and, thus, increases Caspase 3 activity and apoptosis. Along the same lines, APEX1 (1-20) inhibits Caspase 3 cleavage and apoptosis. Furthermore, re-expression of Thioredoxin-1 via lentiviral transduction rescues endothelial cells from APEX1 (21-318)-induced apoptosis. In an in vivo model of restenosis, which is characterized by oxidative stress, endothelial activation, and smooth muscle cell proliferation, Thioredoxin-1 protein levels are reduced in the endothelium of the carotids.

Innovation: APEX1 acts anti-apoptotic in endothelial cells. This anti-apoptotic effect depends on the first 20 amino acids of APEX1.

Conclusion: As proper function of the endothelium during life span is a hallmark for individual health span, a detailed characterization of the functions of the APEX1N-terminus is required to understand all its cellular properties. Antioxid. Redox Signal. 26, 616-629.

Keywords: APEX1; N-terminus; Trx-1; anti-apoptotic.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acids / genetics
  • Amino Acids / metabolism
  • Apoptosis / genetics*
  • Blood Vessels / metabolism
  • Blood Vessels / pathology
  • Carotid Arteries / metabolism
  • Carotid Arteries / pathology
  • Caspase 3 / genetics
  • Caspase 3 / metabolism
  • Cathepsin D / genetics
  • Cell Proliferation / genetics
  • DNA-(Apurinic or Apyrimidinic Site) Lyase / biosynthesis
  • DNA-(Apurinic or Apyrimidinic Site) Lyase / genetics*
  • Endothelial Cells / metabolism
  • Gene Expression Regulation
  • Graft Occlusion, Vascular / genetics*
  • Graft Occlusion, Vascular / pathology
  • Humans
  • Myocytes, Smooth Muscle / metabolism
  • Myocytes, Smooth Muscle / pathology
  • Oxidative Stress / genetics
  • Thioredoxins / biosynthesis*
  • Thioredoxins / genetics

Substances

  • Amino Acids
  • Thioredoxins
  • Caspase 3
  • Cathepsin D
  • APEX1 protein, human
  • DNA-(Apurinic or Apyrimidinic Site) Lyase