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Rictor, an essential component of mTOR complex 2, undergoes caspase-mediated cleavage during apoptosis induced by multiple stimuli

Abstract

Caspase-mediated cleavage of proteins ensures the irreversible commitment of cells to undergo apoptosis, and is thus a hallmark of apoptosis. Rapamycin-insensitive companion of mTOR (rictor) functions primarily as a core and essential component of mTOR complex 2 (mTORC2) to critically regulate cellular homeostasis. However, its role in the regulation of apoptosis is largely unknown. In the current study, we found that rictor was cleaved to generate two small fragments at ~ 50 kD and ~ 130 kD in cells undergoing apoptosis upon treatment with different stimuli such as the death ligand, TRAIL, and the small molecule, AZD9291. This cleavage was abolished when caspases were inhibited and could be reproduced when directly incubating rictor protein and caspase-3 in vitro. Furthermore, the cleavage site of caspase-3 on rictor was mapped at D1244 (VGVD). These findings together robustly demonstrate that rictor is a substrate of caspase-3 and undergoes cleavage during apoptosis. These results add new information for understanding the biology of rictor in the regulation of cell survival and growth.

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All data have been included in this manuscript and materials are available upon request.

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Acknowledgements

We are thankful to Drs. Chunhai Hao and Keqiang Ye for sharing some cell lines and grateful to Dr. Anthea Hammond in our department for editing the manuscript. SY Sun is a Georgia Research Alliance Distinguished Cancer Scientist.

Funding

This study was supported by the Emory Winship Cancer Institute lung cancer research pilot funds (to SYS). S-Y Sun was suported by NIH/NCI R01 CA223220 (to SYS) and R01 CA245386 (to SYS).

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LZ, LZ, YO and SYS conceived and designed the study. LZ, LZ, YO, GQ and ZC performed the experiments and analyzed data. SYS supervised the project, analyzed data and drafted the manuscript. All authors read, edited and approved the manuscript.

Corresponding author

Correspondence to Shi-Yong Sun.

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Zhao, L., Zhu, L., Oh, YT. et al. Rictor, an essential component of mTOR complex 2, undergoes caspase-mediated cleavage during apoptosis induced by multiple stimuli. Apoptosis 26, 338–347 (2021). https://doi.org/10.1007/s10495-021-01676-y

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Keywords

  • Rictor
  • TRAIL
  • EGFR-TKIs
  • Apoptosis