jun-NH2-terminal kinase activation mediated by UV-induced DNA lesions in melanoma and fibroblast cells.
jun-NH2-terminal kinase (JNK) belongs to a family of protein kinases that phosphorylates c-Jun, ATF2, and Elk1 in response to various forms of stress including UV irradiation and heat shock. Although in previous studies we have demonstrated the importance of membrane components for JNK activation by UV irradiation, here we have elucidated the role of DNA damage in this response. We show that in vitro-irradiated or sonicated DNA that is added to proteins prepared from UV-treated cells can further induce JNK activation in a dose-dependent manner. When compared with UV-B (300 nm), UV-C (254 nm), which is better absorbed by the DNA, is significantly more potent in activating JNK. Furthermore, when wavelengths lower than 300 nm were filtered out, UV-B was no longer able to activate JNK. With the aid of melanoma and fibroblast cells, which exhibit different resistances to irradiation and require different UV doses to generate the same number of DNA lesions, we demonstrate that above a threshold level of 0.45 lesions and up to 0.75 lesions per 1875 bp, the degree of JNK activation correlates with the amount of lesions induced by UV-C irradiation. Finally, to explore the role of nuclear and mitochondrial DNA (mtDNA) in mediating JNK activation after UV irradiation, we have used cells that lacks mtDNA. Although the lack of mtDNA did not impair the ability of UV to activate JNK, when enucleated, these cells had lost the ability to activate JNK in response to UV irradiation. Overall, our results suggest that DNA damage in the nuclear compartment is an essential component that acts in concert with membrane-anchored proteins to mediate c-Jun phosphorylation by JNK.
Adler
V
V
Molecular Carcinogenesis Program, American Health Foundation, Valhalla, New York 10595, USA.
Fuchs
S Y
SY
Kim
J
J
Kraft
A
A
King
M P
MP
Pelling
J
J
Ronai
Z
Z
D016475:3T3 Cells / Q000201:enzymology / Q000528:radiation effects; D000818:Animals; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000235:genetics / Q000378:metabolism* / Q000528:radiation effects; D002467:Cell Nucleus / Q000235:genetics / Q000528:radiation effects; D006224:Cricetinae; D004247:DNA / Q000378:metabolism / Q000528:radiation effects; D004249:DNA Damage / Q000502:physiology / Q000528:radiation effects; D005347:Fibroblasts / Q000201:enzymology*; D015870:Gene Expression / Q000528:radiation effects; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism* / Q000528:radiation effects; D048031:JNK Mitogen-Activated Protein Kinases; D008545:Melanoma / Q000201:enzymology*; D051379:Mice; D020928:Mitogen-Activated Protein Kinases; D014466:Ultraviolet Rays
D007527:Isoenzymes; D004247:DNA; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D048031:JNK Mitogen-Activated Protein Kinases; D020928:Mitogen-Activated Protein Kinases