Phosphorylation of OTUB1 at Tyr 26 Stabilizes the MTORC1 Component, Raptor

Overview

Journal Cell Death Differ

Specialty Cell Biology

Date 2022 Aug 4

PMID 35927303

Authors

Seung Un Seo

Seon Min Woo

Min Wook Kim

Eun-Woo Lee

Kyoung-Jin Min

Taeg Kyu Kwon

Affiliations

Soon will be listed here.

Abstract

Raptor plays a critical role in mTORC1 signaling. High expression of Raptor is associated with resistance of cancer cells to PI3K/mTOR inhibitors. Here, we found that OTUB1-stabilized Raptor in a non-canonical manner. Using biochemical assays, we found that the tyrosine 26 residue (Y26) of OTUB1 played a critical role in the interaction between OTUB1 and Raptor. Furthermore, non-receptor tyrosine kinases (Src and SRMS kinases) induced phosphorylation of OTUB1 at Y26, which stabilized Raptor. Interestingly, phosphorylation of OTUB1 at Y26 did not affect the stability of other OTUB1-targeted substrates. However, dephosphorylation of OTUB1 destabilized Raptor and sensitized cancer cells to anti-cancer drugs via mitochondrial reactive oxygen species-mediated mitochondrial dysfunction. Furthermore, we detected high levels of phospho-OTUB1 and Raptor in samples of patients with renal clear carcinoma. Our results suggested that regulation of OTUB1 phosphorylation may be an effective and selective therapeutic target for treating cancers via down-regulation of Raptor.

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References

Edelmann M, Kramer H, Altun M, Kessler B . Post-translational modification of the deubiquitinating enzyme otubain 1 modulates active RhoA levels and susceptibility to Yersinia invasion. FEBS J. 2010; 277(11):2515-30. DOI: 10.1111/j.1742-4658.2010.07665.x. View

Kamba T, McDonald D . Mechanisms of adverse effects of anti-VEGF therapy for cancer. Br J Cancer. 2007; 96(12):1788-95. PMC: 2359962. DOI: 10.1038/sj.bjc.6603813. View

Qayyum T, McArdle P, Lamb G, Jordan F, Orange C, Seywright M . Expression and prognostic significance of Src family members in renal clear cell carcinoma. Br J Cancer. 2012; 107(5):856-63. PMC: 3426751. DOI: 10.1038/bjc.2012.314. View

Wang T, Zhang W, Wang Z, Wu Z, Du B, Li L . RAPTOR promotes colorectal cancer proliferation by inducing mTORC1 and upregulating ribosome assembly factor URB1. Cancer Med. 2019; 9(4):1529-1543. PMC: 7013072. DOI: 10.1002/cam4.2810. View

Xu J, Pham C, Albanese S, Dong Y, Oyama T, Lee C . Mechanistically distinct cancer-associated mTOR activation clusters predict sensitivity to rapamycin. J Clin Invest. 2016; 126(9):3526-40. PMC: 5004947. DOI: 10.1172/JCI86120. View

Kohmura N, Yagi T, Tomooka Y, Oyanagi M, Kominami R, Takeda N . A novel nonreceptor tyrosine kinase, Srm: cloning and targeted disruption. Mol Cell Biol. 1994; 14(10):6915-25. PMC: 359222. DOI: 10.1128/mcb.14.10.6915-6925.1994. View

Pasupala N, Morrow M, Que L, Malynn B, Ma A, Wolberger C . OTUB1 non-catalytically stabilizes the E2 ubiquitin-conjugating enzyme UBE2E1 by preventing its autoubiquitination. J Biol Chem. 2018; 293(47):18285-18295. PMC: 6254341. DOI: 10.1074/jbc.RA118.004677. View

Wiener R, Zhang X, Wang T, Wolberger C . The mechanism of OTUB1-mediated inhibition of ubiquitination. Nature. 2012; 483(7391):618-22. PMC: 3319311. DOI: 10.1038/nature10911. View

Liu T, Jiang L, Tavana O, Gu W . The Deubiquitylase OTUB1 Mediates Ferroptosis via Stabilization of SLC7A11. Cancer Res. 2019; 79(8):1913-1924. PMC: 6467774. DOI: 10.1158/0008-5472.CAN-18-3037. View

10.

Park J, Yang S, Zhuang W, Bera A, Liu Y, Gurbani D . The nonreceptor tyrosine kinase SRMS inhibits autophagy and promotes tumor growth by phosphorylating the scaffolding protein FKBP51. PLoS Biol. 2021; 19(6):e3001281. PMC: 8202955. DOI: 10.1371/journal.pbio.3001281. View

11.

Cho D, Cohen M, Panka D, Collins M, Ghebremichael M, Atkins M . The efficacy of the novel dual PI3-kinase/mTOR inhibitor NVP-BEZ235 compared with rapamycin in renal cell carcinoma. Clin Cancer Res. 2010; 16(14):3628-38. PMC: 2905505. DOI: 10.1158/1078-0432.CCR-09-3022. View

12.

Zhou Y, Wu J, Fu X, Du W, Zhou L, Meng X . OTUB1 promotes metastasis and serves as a marker of poor prognosis in colorectal cancer. Mol Cancer. 2014; 13:258. PMC: 4351937. DOI: 10.1186/1476-4598-13-258. View

13.

Yusenko M, Kuiper R, Boethe T, Ljungberg B, Geurts van Kessel A, Kovacs G . High-resolution DNA copy number and gene expression analyses distinguish chromophobe renal cell carcinomas and renal oncocytomas. BMC Cancer. 2009; 9:152. PMC: 2686725. DOI: 10.1186/1471-2407-9-152. View

14.

Herhaus L, Perez-Oliva A, Cozza G, Gourlay R, Weidlich S, Campbell D . Casein kinase 2 (CK2) phosphorylates the deubiquitylase OTUB1 at Ser16 to trigger its nuclear localization. Sci Signal. 2015; 8(372):ra35. PMC: 4421874. DOI: 10.1126/scisignal.aaa0441. View

15.

Bridges C, Tan M, Premarathne S, Nanayakkara D, Bellette B, Zencak D . USP9X deubiquitylating enzyme maintains RAPTOR protein levels, mTORC1 signalling and proliferation in neural progenitors. Sci Rep. 2017; 7(1):391. PMC: 5427856. DOI: 10.1038/s41598-017-00149-0. View

16.

Zhao L, Wang X, Yu Y, Deng L, Chen L, Peng X . OTUB1 protein suppresses mTOR complex 1 (mTORC1) activity by deubiquitinating the mTORC1 inhibitor DEPTOR. J Biol Chem. 2018; 293(13):4883-4892. PMC: 5880136. DOI: 10.1074/jbc.M117.809533. View

17.

Seo J, Lee E, Shin J, Seong D, Nam Y, Jeong M . K6 linked polyubiquitylation of FADD by CHIP prevents death inducing signaling complex formation suppressing cell death. Oncogene. 2018; 37(36):4994-5006. DOI: 10.1038/s41388-018-0323-z. View

18.

Zhou H, Liu Y, Zhu R, Ding F, Cao X, Lin D . OTUB1 promotes esophageal squamous cell carcinoma metastasis through modulating Snail stability. Oncogene. 2018; 37(25):3356-3368. DOI: 10.1038/s41388-018-0224-1. View

19.

Goncharov T, Niessen K, de Almagro M, Izrael-Tomasevic A, Fedorova A, Varfolomeev E . OTUB1 modulates c-IAP1 stability to regulate signalling pathways. EMBO J. 2013; 32(8):1103-14. PMC: 3630360. DOI: 10.1038/emboj.2013.62. View

20.

Vojtechova M, Tureckova J, Kucerova D, Sloncova E, Vachtenheim J, Tuhackova Z . Regulation of mTORC1 signaling by Src kinase activity is Akt1-independent in RSV-transformed cells. Neoplasia. 2008; 10(2):99-107. PMC: 2244684. DOI: 10.1593/neo.07905. View