Thioredoxin-1 maintains mTOR function during oxidative stress in cardiomyocytes.

5 years ago

Thioredoxin-1 maintains mTOR function during oxidative stress in cardiomyocytes.

Sciarretta, Yaginuma, Chin, Hirata, Oka, Yodoi, Chen, Schesing, Shao, Zhai, Suzuki, Bhat, Hirabayashi, Nagarajan, Naito, Saito, Sadoshima

Thioredoxin 1 (Trx1) is a 12 kDa oxidoreductase that catalyzes thiol-disulfide exchange reactions to reduce proteins with disulfide bonds. As such, Trx1 helps protect the heart against stresses, such as ischemia and pressure overload. Mechanistic target of rapamycin (mTOR) is a serine/threonine kinase that regulates cell growth, metabolism and survival. We have previously shown that mTOR activity is increased in response to myocardial ischemia-reperfusion injury. However, whether Trx1 interacts with mTOR to preserve heart function remains unknown. Using a substrate-trapping mutant of Trx1 (Trx1C35S), we here show that mTOR is a direct interacting partner of Trx1 in the heart. In response to H2O2 treatment in cardiomyocytes, mTOR exhibited a high molecular weight shift in non-reducing SDS-PAGE in a 2-mercaptoethanol sensitive manner, suggesting that mTOR is oxidized and forms disulfide bonds with itself or other proteins. The mTOR oxidation was accompanied by reduced phosphorylation of endogenous substrates, such as S6 kinase (S6K) and 4E-Binding Protein 1 (4EBP1) in cardiomyocytes. Immune complex kinase assays disclosed that the H2O2 treatment diminished mTOR kinase activity, indicating that mTOR is inhibited by oxidation. Of note, Trx1 overexpression attenuated both H2O2-mediated mTOR oxidation and inhibition, whereas Trx1 knockdown increased mTOR oxidation and inhibition. Moreover, Trx1 normalized H2O2- induced downregulation of metabolic genes and stimulation of cell death, and an mTOR inhibitor abolished Trx1-mediated rescue of gene expression. H2O2-induced oxidation and inhibition of mTOR were attenuated when Cys1483 of mTOR was mutated to phenylalanine. These results suggest that Trx1 protects cardiomyocytes against stress by reducing mTOR at Cys1483, thereby preserving the activity of mTOR and inhibiting cell death.

Publisher URL: http://doi.org/10.1074/jbc.M117.807735

DOI: 10.1074/jbc.M117.807735