Glyceraldehyde-3-phosphate dehydrogenase regulates activation of c-Jun N-terminal kinase under oxidative stress

• Published in: Biochemical and biophysical research communications Vol. 657; pp. 1 - 7
• Main Authors: Itakura, Masanori, Kubo, Takeya, Kaneshige, Akihiro, Nakajima, Hidemitsu
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 21.05.2023
• Subjects: c-Jun N-terminal kinase (JNK); Cysteine - metabolism; Glyceraldehyde-3-phosphate dehydrogenase (GAPDH); Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating) - metabolism; H2O2; HEK293 Cells; Humans; Hydrogen Peroxide - pharmacology; JNK Mitogen-Activated Protein Kinases - metabolism; Mitogen-Activated Protein Kinase 8 - pharmacology; Mitogen-activated protein kinases (MAPKs); Oxidative Stress; p38 Mitogen-Activated Protein Kinases - metabolism; c-Jun N-terminal kinase (JNK); Oxidative stress; Mitogen-activated protein kinases (MAPKs); Glyceraldehyde-3-phosphate dehydrogenase (GAPDH); H2O2; HO
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1016/j.bbrc.2023.03.044

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) acts as a sensor under oxidative stress, leading to induction of various biological responses. Given that mitogen-activated protein kinase (MAPK) signaling pathways mediate cellular responses to a wide variety of stimuli, including oxidative stress, here, we aimed to elucidate whether a cross-talk cascade between GAPDH and MAPKs occurs under oxidative stress. Of the three typical MAPKs investigated—extracellular signal-regulated kinase, p38, and c-Jun N-terminal kinase (JNK)—we found that hydrogen peroxide (H2O2)-induced JNK activation is significantly reduced in HEK293 cells treated with small-interfering (si)RNA targeting GAPDH. Co-immunoprecipitation with a GAPDH antibody further revealed protein–protein interactions between GAPDH and JNK in H2O2-stmulated cells. Notably, both JNK activation and these interactions depend on oxidation of the active-site cysteine (Cys152) in GAPDH, as demonstrated by rescue experiments with either exogenous wild-type GAPDH or the cysteine-substituted mutant (C152A) in endogenous GAPDH-knockdown HEK293 cells. Moreover, H2O2-induced translocation of Bcl-2-associated X protein (Bax) into mitochondria, which occurs downstream of JNK activation, is attenuated by endogenous GAPDH knockdown in HEK293 cells. These results suggest a novel role for GAPDH in the JNK signaling pathway under oxidative stress. •GAPDH is involved in H2O2-induced JNK activation.•GAPDH Cys152 plays a critical role in JNK activation via protein–protein interactions.•GAPDH induces Bax mitochondrial translocation following JNK activation.•Our findings uncover an additional non-glycolytic function for GAPDH.