Exp Mol Med.  2008 Jun;40(3):276-285. 10.3858/emm.2008.40.3.276.

Cytosolic accumulation of gammaH2AX is associated with tropomyosin-related kinase A-induced cell death in U2OS cells

• Jung EJ
• Kim CW
• Kim DR

• KMID: 756329
• DOI: http://doi.org/10.3858/emm.2008.40.3.276

Abstract

Tropomyosin-related kinase A (TrkA) plays an important role in cell survival, differentiation, and apoptosis in various neuronal and nonneuronal cell types. Here we show that TrkA overexpression by the Tet-On system mimics NGF-mediated activation pathways in the absence of nerve growth factor (NGF) stimulation in U2OS cells. In addition, p53 upregulation upon DNA damage was inhibited by TrkA, and p21 was upregulated by TrkA in a p53-independent manner. TrkA overexpression caused cell death by interrupting cell cycle progression, and TrkA-induced cell death was diminished in the presence of its specific inhibitor GW441756. Interestingly, TrkA-mediated cell death was strongly related to gammaH2AX production and poly (ADP-ribose) polymerase cleavage in the absence of DNA damage inducer. In this study, we also reveal thatgammagammaH2AX production by TrkA is blocked by TrkA kinase inhibitors K-252a and GW441756, and it is also significantly inhibited by JNK inhibitor SP600125. Moreover, reduction of cell viability by TrkA was strongly suppressed by SP600125 treatment, suggesting a critical role of JNK in TrkA-induced cell death. We also found that gammaH2AX and TrkA were colocalized in cytosol in the absence of DNA damage, and the nuclear localization of gammaH2AX induced by DNA damage was partly altered to cytosol by TrkA overexpression. Our results suggest that the abnormal cytosolic accumulation of gammaH2AX is implicated in TrkA-induced cell death in the absence of DNA damage.

Keyword

cell death; DNA damage; H2AFX protein, human; JNK mitogen-activated protein kinases; receptor, trkA

MeSH Terms

Anthracenes/pharmacology
Apoptosis/drug effects/*genetics
Carbazoles/pharmacology
Cell Cycle/drug effects/genetics
Cell Line, Tumor
Cyclin-Dependent Kinase Inhibitor p21/*biosynthesis/genetics
Cytosol/drug effects/enzymology/ultrastructure
DNA Damage/drug effects/genetics
Doxorubicin/pharmacology
Histones/*metabolism
Humans
Indole Alkaloids/pharmacology
MAP Kinase Kinase 4/antagonists & inhibitors
Nerve Growth Factor/antagonists & inhibitors/metabolism
Phosphorylation/drug effects
Protein Binding
*Protein Transport/drug effects/genetics
Receptor, trkA/antagonists & inhibitors/*genetics/metabolism
Signal Transduction
Transfection