NOX2, p22phox and p47phox are targeted to the nuclear pore complex in ischemic cardiomyocytes colocalizing with local reactive oxygen species
Background: NADPH oxidases play an essential role in reactive oxygen species (ROS)-based signaling in the heart. Previously, we have demonstrated that (peri)nuclear expression of the catalytic NADPH oxidase subunit NOX2 in stressed cardiomyocytes, e.g. under ischemia or high concentrations of homocysteine, is an important step in the induction of apoptosis in these cells. Here this ischemia-induced nuclear targeting and activation of NOX2 was specified in cardiomyocytes. Methods: The effect of ischemia, mimicked by metabolic inhibition, on nuclear localization of NOX2 and the NADPH oxidase subunits p22phoxand p47phox, was analyzed in rat neonatal cardiomyoblasts (H9c2 cells) using Western blot, immuno-electron microscopy and digital-imaging microscopy. Results: NOX2 expression significantly increased in nuclear fractions of ischemic H9c2 cells. In addition, in these cells NOX2 was found to colocalize in the nuclear envelope with nuclear pore complexes, p22phox, p47phoxand nitrotyrosine residues, a marker for the generation of ROS. Inhibition of NADPH oxidase activity, with apocynin and DPI, significantly reduced (peri)nuclear expression of nitrotyrosine. Conclusion: We for the first time show that NOX2, p22phoxand p47phoxare targeted to and produce ROS at the nuclear pore complex in ischemic cardiomyocytes.
|Keywords||Apoptosis, Cardiomyocytes, NADPH oxidase, Nuclear pore complex, Reactive oxygen species|
|Persistent URL||dx.doi.org/10.1159/000329968, hdl.handle.net/1765/34195|
Hahn, N.E., Meischl, C., Wijnker, P.J.M., Musters, R.J.P., Fornerod, M.W.J., Janssen, H.W.R.M., … van Rossum, A.C.. (2011). NOX2, p22phox and p47phox are targeted to the nuclear pore complex in ischemic cardiomyocytes colocalizing with local reactive oxygen species. Cellular Physiology and Biochemistry, 27(5), 471–478. doi:10.1159/000329968