Identification of a hydrogen peroxide-induced PP1-JNK1-Sp1 signaling pathway for gene regulation.
Oxidative stress often results in changes in gene expression through the regulation of transcription factors. In this study, we examine how Sp1 phosphorylation is regulated by H(2)O(2) in a human alveolar epithelial cell line (HAE). Treatment of HAE cells with H(2)O(2) increases phosphorylation of Sp1 and activates JNK. To establish a relationship between JNK and Sp1, we show that JNK activator anisomycin increases Sp1 phosphorylation, and JNK inhibitors as well as dominant-negative JNK1 attenuate H(2)O(2)-induced Sp1 phosphorylation. Additionally, JNK1 directly phosphorylates Sp1 in vitro, reducing Sp1 binding to DNA. These results demonstrate the role of JNK in H(2)O(2)-induced Sp1 phosphorylation. Because H(2)O(2) inhibits Ser/Thr protein phosphatase-1 (PP1), we examined the role of PP1 in the regulation of JNK. Similar to H(2)O(2), inhibition of PP1 induces phosphorylation of Sp1 and activation of JNK in HAE cells. Inhibition of JNK activity using either inhibitors or dominant-negative mutant JNK1 suppresses PP1 inhibition-induced Sp1 phosphorylation. Furthermore, PP1 directly inactivates JNK1 in vitro. These data suggest that 1) H(2)O(2) increases the phosphorylation level of Sp1, 2) Sp1 is a target of the JNK pathway, 3) PP1 regulates JNK activation, and 4) the "PP1-JNK" pathway plays a role in H(2)O(2)-induced Sp1 phosphorylation in lung epithelial cells.
Chu
Shijian
S
McGuire Veterans Affairs Medical Center, 1201 Broad Rock Blvd., Richmond, VA 23249, USA. schu@hsc.vcu.edu
Ferro
Thomas J
TJ
D000328:Adult; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000502:physiology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects / Q000502:physiology; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D048055:Mitogen-Activated Protein Kinase 8 / Q000378:metabolism*; D016877:Oxidants / Q000494:pharmacology; D018384:Oxidative Stress / Q000187:drug effects / Q000502:physiology; D010749:Phosphoprotein Phosphatases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D010766:Phosphorylation / Q000187:drug effects; D054645:Protein Phosphatase 1; D011650:Pulmonary Alveoli / Q000166:cytology / Q000378:metabolism*; D020545:Respiratory Mucosa / Q000166:cytology / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology; D016329:Sp1 Transcription Factor / Q000378:metabolism*
D004791:Enzyme Inhibitors; D016877:Oxidants; D016329:Sp1 Transcription Factor; D006861:Hydrogen Peroxide; D048055:Mitogen-Activated Protein Kinase 8; D010749:Phosphoprotein Phosphatases; D054645:Protein Phosphatase 1