Nucleic Acids Research, 2001, Vol. 29, No. 14 3116-3122
© 2001 Oxford University Press
Activation of APE/Ref-1 redox activity is mediated by reactive oxygen species and PKC phosphorylation
DNA Damage and Repair Laboratory, Pennington Biomedical Research Center, Louisiana State University, 6400 Perkins, Baton Rouge, LA 70808, USA and 1Department of Pediatrics and Biochemistry and Molecular Biology, Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, IN 46202, USA
Reactive oxygen species (ROS) arise through normal cellular aerobic respiration, and, in combination with external sources such as ionizing radiation, cigarette tar and smoke, and particulate matter generated by combustion, can have a profound negative effect on cellular macromolecules such as DNA that may lead to a number of human pathological disorders including accelerated aging and cancer. A major end product of ROS damage to DNA is the formation of apurinic/apyrimidinic (AP) sites, which without removal are known to halt mRNA and DNA synthesis, or act as non-coding lesions resulting in the increased generation of DNA mutations. In human cells, the major enzyme in correcting the deleterious effects of AP sites in DNA is through the participation of AP endonuclease (APE), which initiates the removal of baseless sites in DNA through the catalytic scission of the phosphodiester bond 5' and adjacent to an AP site. Interestingly, APE also possesses an activity (Ref-1) that controls the redox status of a number of transcription factors including Fos and Jun. The means by which APE/Ref-1 is directed to carry out such disparate roles are unknown. The presence of a number of phosphorylation sites scattered throughout both functional domains of APE/Ref-1 however offered one possible mechanism that we reasoned could play a role in dictating how this protein responds to different stimuli. Here we show that the in vitro redox activity of APE/Ref-1 is stimulated by PKC phosphorylation. Furthermore, when human cells were exposed to the PKC activator phorbol 12-myristate 13-acetate, an increase in redox activity was observed that corresponded to an increase in the phosphorylation status of APE/Ref-1. Importantly, human cells exposed to the oxidizing agent hypochlorite, followed by methyl methanesulfanate, responded with an increase in redox activity by APE/Ref-1 that also involved an increase in PKC activity and a corresponding increase in the phosphorylation of APE/Ref-1. These results suggest that the ability of APE/Ref-1 to perform its in vivo redox function is correlated to its susceptibility to PKC phosphorylation that notably occurs in response to DNA damaging agents.
* To whom correspondence should be addressed. Tel: +1 225 763 0937; Fax: +1 225 763 3030; Email: deutscwa{at}pbrc.edu
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  H. Fung, P. Liu, and B. Demple ATF4-Dependent Oxidative Induction of the DNA Repair Enzyme Ape1 Counteracts Arsenite Cytotoxicity and Suppresses Arsenite-Mediated Mutagenesis Mol. Cell. Biol., December 15, 2007; 27(24): 8834 - 8847. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. El-Mouatassim, S. Bilotto, G. L. Russo, E. Tosti, and Y. Menezo APEX/Ref-1 (apurinic/apyrimidic endonuclease DNA-repair gene) expression in human and ascidian (Ciona intestinalis) gametes and embryos Mol. Hum. Reprod., August 1, 2007; 13(8): 549 - 556. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 I.-Y. Chang, S.-H. Kim, H.-J. Cho, D. Y. Lee, M.-H. Kim, M.-H. Chung, and H. J. You Human AP endonuclease suppresses DNA mismatch repair activity leading to microsatellite instability Nucleic Acids Res., September 7, 2005; 33(16): 5073 - 5081. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Pines, L. Perrone, N. Bivi, M. Romanello, G. Damante, M. Gulisano, M. R. Kelley, F. Quadrifoglio, and G. Tell Activation of APE1/Ref-1 is dependent on reactive oxygen species generated after purinergic receptor stimulation by ATP Nucleic Acids Res., August 2, 2005; 33(14): 4379 - 4394. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. R. Trzeciak, S. G. Nyaga, P. Jaruga, A. Lohani, M. Dizdaroglu, and M. K. Evans Cellular repair of oxidatively induced DNA base lesions is defective in prostate cancer cell lines, PC-3 and DU-145 Carcinogenesis, August 1, 2004; 25(8): 1359 - 1370. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. Hedley, M. Pintilie, J. Woo, T. Nicklee, A. Morrison, D. Birle, A. Fyles, M. Milosevic, and R. Hill Up-Regulation of the Redox Mediators Thioredoxin and Apurinic/Apyrimidinic Excision (APE)/Ref-1 in Hypoxic Microregions of Invasive Cervical Carcinomas, Mapped Using Multispectral, Wide-Field Fluorescence Image Analysis Am. J. Pathol., February 1, 2004; 164(2): 557 - 565. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. Upadhyay and D. W. Kamp Asbestos-Induced Pulmonary Toxicity: Role of DNA Damage and Apoptosis Experimental Biology and Medicine, June 1, 2003; 228(6): 650 - 659. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
F. Dantzer, L. Luna, M. Bjoras, and E. Seeberg Human OGG1 undergoes serine phosphorylation and associates with the nuclear matrix and mitotic chromatin in vivo Nucleic Acids Res., June 1, 2002; 30(11): 2349 - 2357. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. M. Flaherty, M. M. Monick, and G. W. Hunninghake AP Endonucleases and the Many Functions of Ref-1 Am. J. Respir. Cell Mol. Biol., December 1, 2001; 25(6): 664 - 667. [Full Text] [PDF]
|
|