Reactive oxygen species- and dimerization-induced activation of apoptosis signal-regulating kinase 1 in tumor necrosis factor-alpha signal transduction.
Reactive oxygen species (ROS) have been implicated in the induction of apoptosis by tumor necrosis factor-alpha (TNFalpha) and other cytotoxic insults, although the molecule(s) regulated by ROS in TNFalpha signaling have not been identified. Apoptosis signal-regulating kinase 1 (ASK1) is a member of the mitogen-activated protein kinase kinase kinase (MAPKKK) superfamily that has been shown to be activated during TNFalpha-induced apoptosis. ASK1 increases apoptosis when overexpressed, but the mechanism of ASK1 activation and the mechanisms of ASK1-induced apoptosis are unclear. We now report that hydrogen peroxide induces the activation of ASK1 in 293 cells. TNFalpha-induced activation of ASK1 was inhibited by antioxidants. Hydrogen peroxide-induced apoptosis was markedly enhanced by the expression of ASK1. These results suggest that TNFalpha-induced activation of ASK1 is mediated by ROS. We also examined how ASK1 activity is regulated by ROS. We found that ASK1 formed dimers or higher order oligomers in 293 cells. TNFalpha or hydrogen peroxide treatment increased the dimeric form of ASK1, and pretreatment with N-acetylcysteine decreased it. Furthermore, synthetic dimerization of an ASK1-gyrase B fusion protein by coumermycin resulted in substantial activation of ASK1, suggesting that dimerization of ASK1 is sufficient for its activation. These results taken together suggest that TNFalpha causes ASK1 activation via ROS-mediated dimerization of ASK1.
Gotoh
Y
Y
Fred Hutchinson Cancer Research Center, A2-025, Seattle, Washington 98109, USA.
Cooper
J A
JA
D001483:Base Sequence; D002460:Cell Line; D027081:DNA Gyrase; D017931:DNA Primers; D004250:DNA Topoisomerases, Type II / Q000378:metabolism; D019281:Dimerization; D004789:Enzyme Activation; D006801:Humans; D020930:MAP Kinase Kinase Kinases; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D017382:Reactive Oxygen Species; D015398:Signal Transduction / Q000187:drug effects*; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*
D017931:DNA Primers; D017382:Reactive Oxygen Species; D014409:Tumor Necrosis Factor-alpha; D017346:Protein Serine-Threonine Kinases; D020930:MAP Kinase Kinase Kinases; D027081:DNA Gyrase; D004250:DNA Topoisomerases, Type II