NOX4 activity is determined by mRNA levels and reveals a unique pattern of ROS generation

We report the generation of a cell line that expresses NOX4 upon tet induction

Lena Serrander; Laetitia Cartier; Karen Bedard; Botond Banfi; Bernard Lardy; Olivier Plastre; Andrzej Sienkiewicz; Lászlo Fórró; Werner Schlegel; Karl-Heinz Krause

2007

Scholarcy highlights

  • NOX4 is a member of the NOX family of NADPH oxidases
  • Cells were induced with tet concentrations ranging from 1.25 fg/ml to 10 μg/ml, and NOX4 activity was assessed by measuring reactive oxygen species generation
  • Given the fact that NOX4 generates substantial amounts of ROS without a need for cell stimulation, a stable expression system has inherent dangers: damage to DNA which could lead to cellular senescence or, in cells with deficient p53 function, to an accumulation of DNA mutations; damage to mitochondria which may lead to increased mitochondrial ROS generation; and permanent upregulation of antioxidant-responsive genes, which would make it difficult to distinguish the acute NOX4 effects from a longterm antioxidant response
  • One of the striking results of our study is that NOX4 mRNA levels are closely followed by NOX4 activity
  • Our results based on NOX4 function suggest that the NOX4 protein is rapidly synthesized and degraded
  • A heterologous expression system cannot determine whether NOX4 is regulated on a transcriptional level in endogenously NOX4-expressing cells
  • 10 μM diphenyleneiodonium chloride ; no reactive oxygen species generation was detected unless NOX4 expression was induced by tet

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