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Chuck Reynolds
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What does Resveratrol induce.? P-7

Published on 6/15/2019
For additional information  Click Here

Resveratrol induces autophagy by directly inhibiting mTOR through ATP competition P-7

mTOR-associated DEPTOR level is not changed by resveratrol

DEPTOR is a negative regulator of both mTORC1 and mTORC2. Liu et al. showed that resveratrol increased the interaction affinity between mTOR and DEPTOR and suggested that DEPTOR is required for the suppression of mTOR activity upon resveratrol treatment21. To examine the involvement of DEPTOR, we conducted a pull-down assay of FLAG-tagged mTOR under various conditions, including resveratrol treatment.

Rapamycin was used as a positive control for chemically-induced complex dissociation. Unexpectedly, in this condition, both the 10 min and 60 min resveratrol treatments did not increase the affinity between mTOR and DEPTOR although resveratrol clearly decreased mTOR activity. Treatment with rapamycin led to the detachment of Raptor from mTOR.

Resveratrol inhibits mTOR kinase activity in cell-free systems

Next, we examined whether resveratrol directly suppresses mTOR activity. To address this question, we performed an in vitro kinase assay. We observed that resveratrol dose-dependently decreased the phosphorylation level of 4E-BP1.. In addition to the experiments with full-length mTOR, we used a recombinant fragment mTOR that some regulatory domains are deleted, but still contains the intact kinase domain for a kinase assay.

Notably, resveratrol could decrease the phosphorylation of S6K, a substrate of mTOR, similar to the results of immunoprecipitated full-length mTOR . Additionally, we found that resveratrol has an IC50 value of ~10 μM against mTOR kinase activity. Taken together, resveratrol inhibits mTOR kinase activity in cell-free systems.

Article Produced By
Department of Life Sciences, Pohang University of Science and Technology, Pohang 790-784, Republic of Korea
School of Interdisciplinary Bioscience and Bioengineering, Pohang University of Science and Technology, Pohang 790-784, Republic of Korea
School of Life Sciences, Ulsan National Institute of Science and Technology, Ulsan 689-798, Republic of Korea
Department of Bioscience and Biotechnology, Sejong University, 98 Kunja-Dong, Kwangjin-Ku, Seoul 143-747, Republic of Korea

(All these Articles can be read with this URL: )

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