MARCH1 Negatively Regulates TBK1-mTOR Signaling Pathway by Ubiquitinating TBK1

Overview

Journal BMC Cancer

Publisher Biomed Central

Specialty Oncology

Date 2024 Jul 26

PMID 39061024

Authors

Xiao Li

Kai Cheng

Meng-Di Shang

Yong Yang

Bin Hu

Xi Wang

Xiao-Dan Wei

Yan-Chun Han

Xiao-Gang Zhang

Meng-Hua Dong

Zhen-Lin Yang

Jiu-Qiang Wang

Affiliations

Soon will be listed here.

Abstract

Background: TBK1 positively regulates the growth factor-mediated mTOR signaling pathway by phosphorylating mTOR. However, it remains unclear how the TBK1-mTOR signaling pathway is regulated. Considering that STING not only interacts with TBK1 but also with MARCH1, we speculated that MARCH1 might regulate the mTOR signaling pathway by targeting TBK1. The aim of this study was to determine whether MARCH1 regulates the mTOR signaling pathway by targeting TBK1.

Methods: The co-immunoprecipitation (Co-IP) assay was used to verify the interaction between MARCH1 with STING or TBK1. The ubiquitination of STING or TBK1 was analyzed using denatured co-immunoprecipitation. The level of proteins detected in the co-immunoprecipitation or denatured co-immunoprecipitation samples were determined by Western blotting. Stable knocked-down cells were constructed by infecting lentivirus bearing the related shRNA sequences. Scratch wound healing and clonogenic cell survival assays were used to detect the migration and proliferation of breast cancer cells.

Results: We showed that MARCH1 played an important role in growth factor-induced the TBK1- mTOR signaling pathway. MARCH1 overexpression attenuated the growth factor-induced activation of mTOR signaling pathway, whereas its deficiency resulted in the opposite effect. Mechanistically, MARCH1 interacted with and promoted the K63-linked ubiquitination of TBK1. This ubiquitination of TBK1 then attenuated its interaction with mTOR, thereby inhibiting the growth factor-induced mTOR signaling pathway. Importantly, faster proliferation induced by MARCH1 deficiency was weakened by mTOR, STING, or TBK1 inhibition.

Conclusion: MARCH1 suppressed growth factors mediated the mTOR signaling pathway by targeting the STING-TBK1-mTOR axis.

References

Corrales L, Glickman L, McWhirter S, Kanne D, Sivick K, Katibah G . Direct Activation of STING in the Tumor Microenvironment Leads to Potent and Systemic Tumor Regression and Immunity. Cell Rep. 2015; 11(7):1018-30. PMC: 4440852. DOI: 10.1016/j.celrep.2015.04.031. View

Jacinto E, Loewith R, Schmidt A, Lin S, Ruegg M, Hall A . Mammalian TOR complex 2 controls the actin cytoskeleton and is rapamycin insensitive. Nat Cell Biol. 2004; 6(11):1122-8. DOI: 10.1038/ncb1183. View

Galluzzi L, Vanpouille-Box C, Bakhoum S, Demaria S . SnapShot: CGAS-STING Signaling. Cell. 2018; 173(1):276-276.e1. DOI: 10.1016/j.cell.2018.03.015. View

Woo S, Fuertes M, Corrales L, Spranger S, Furdyna M, Leung M . STING-dependent cytosolic DNA sensing mediates innate immune recognition of immunogenic tumors. Immunity. 2014; 41(5):830-42. PMC: 4384884. DOI: 10.1016/j.immuni.2014.10.017. View

Baravalle G, Park H, McSweeney M, Ohmura-Hoshino M, Matsuki Y, Ishido S . Ubiquitination of CD86 is a key mechanism in regulating antigen presentation by dendritic cells. J Immunol. 2011; 187(6):2966-73. PMC: 4496154. DOI: 10.4049/jimmunol.1101643. View

Clark K, Peggie M, Plater L, Sorcek R, Young E, Madwed J . Novel cross-talk within the IKK family controls innate immunity. Biochem J. 2010; 434(1):93-104. DOI: 10.1042/BJ20101701. View

Sarbassov D, Guertin D, Ali S, Sabatini D . Phosphorylation and regulation of Akt/PKB by the rictor-mTOR complex. Science. 2005; 307(5712):1098-101. DOI: 10.1126/science.1106148. View

Wang C, Wang X, Veleeparambil M, Kessler P, Willard B, Chattopadhyay S . EGFR-mediated tyrosine phosphorylation of STING determines its trafficking route and cellular innate immunity functions. EMBO J. 2020; 39(22):e104106. PMC: 7667877. DOI: 10.15252/embj.2019104106. View

Xie X, Zhang D, Zhao B, Lu M, You M, Condorelli G . IkappaB kinase epsilon and TANK-binding kinase 1 activate AKT by direct phosphorylation. Proc Natl Acad Sci U S A. 2011; 108(16):6474-9. PMC: 3081021. DOI: 10.1073/pnas.1016132108. View

10.

Jiang Q, Guan Y, Zheng J, Lu H . TBK1 promotes thyroid cancer progress by activating the PI3K/Akt/mTOR signaling pathway. Immun Inflamm Dis. 2023; 11(3):e796. PMC: 10013413. DOI: 10.1002/iid3.796. View

11.

Lin H, Li S, Shu H . The Membrane-Associated MARCH E3 Ligase Family: Emerging Roles in Immune Regulation. Front Immunol. 2019; 10:1751. PMC: 6669941. DOI: 10.3389/fimmu.2019.01751. View

12.

Reus J, Trivino-Soto G, Wu L, Kokott K, Lim E . SV40 Large T Antigen Is Not Responsible for the Loss of STING in 293T Cells but Can Inhibit cGAS-STING Interferon Induction. Viruses. 2020; 12(2). PMC: 7077178. DOI: 10.3390/v12020137. View

13.

Vasiyani H, Mane M, Rana K, Shinde A, Roy M, Singh J . DNA damage induces STING mediated IL-6-STAT3 survival pathway in triple-negative breast cancer cells and decreased survival of breast cancer patients. Apoptosis. 2022; 27(11-12):961-978. DOI: 10.1007/s10495-022-01763-8. View

14.

Jacinto E, Facchinetti V, Liu D, Soto N, Wei S, Jung S . SIN1/MIP1 maintains rictor-mTOR complex integrity and regulates Akt phosphorylation and substrate specificity. Cell. 2006; 127(1):125-37. DOI: 10.1016/j.cell.2006.08.033. View

15.

Nazio F, Strappazzon F, Antonioli M, Bielli P, Cianfanelli V, Bordi M . mTOR inhibits autophagy by controlling ULK1 ubiquitylation, self-association and function through AMBRA1 and TRAF6. Nat Cell Biol. 2013; 15(4):406-16. DOI: 10.1038/ncb2708. View

16.

Xiao Y, Zou Q, Xie X, Liu T, Li H, Jie Z . The kinase TBK1 functions in dendritic cells to regulate T cell homeostasis, autoimmunity, and antitumor immunity. J Exp Med. 2017; 214(5):1493-1507. PMC: 5413337. DOI: 10.1084/jem.20161524. View

17.

Xu Z, Liu J, Liu Z, Zhang H . MARCH1 as a novel immune-related prognostic biomarker that shapes an inflamed tumor microenvironment in lung adenocarcinoma. Front Oncol. 2022; 12:1008753. PMC: 9606618. DOI: 10.3389/fonc.2022.1008753. View

18.

Su Y, Feng W, Zhong G, Ya Y, Du Z, Shi J . ciRs-6 upregulates March1 to suppress bladder cancer growth by sponging miR-653. Aging (Albany NY). 2019; 11(23):11202-11223. PMC: 6932879. DOI: 10.18632/aging.102525. View

19.

Tang C, Zundell J, Ranatunga S, Lin C, Nefedova Y, Del Valle J . Agonist-Mediated Activation of STING Induces Apoptosis in Malignant B Cells. Cancer Res. 2016; 76(8):2137-52. PMC: 4873432. DOI: 10.1158/0008-5472.CAN-15-1885. View

20.

Ye M, Hu Y, Zhao B, Mou Q, Ni Y, Luo J . TBK1 Knockdown Alleviates Axonal Transport Deficits in Retinal Ganglion Cells Via mTORC1 Activation in a Retinal Damage Mouse Model. Invest Ophthalmol Vis Sci. 2023; 64(10):1. PMC: 10324417. DOI: 10.1167/iovs.64.10.1. View