PP2A-dependent TFEB Activation is Blocked by PIKfyve-induced MTORC1 Activity

Overview

Journal Mol Biol Cell

Specialties Cell Biology
Molecular Biology

Date 2022 Jan 12

PMID 35020443

Authors

Junya Hasegawa

Emi Tokuda

Yao Yao

Takehiko Sasaki

Ken Inoki

Lois S Weisman

Affiliations

Soon will be listed here.

Abstract

Transcriptional factor EB (TFEB) is a master regulator of genes required for autophagy and lysosomal function. The nuclear localization of TFEB is blocked by the mechanistic target of rapamycin complex 1 (mTORC1)-dependent phosphorylation of TFEB at multiple sites including Ser-211. Here we show that inhibition of PIKfyve, which produces phosphatidylinositol 3,5-bisphosphate on endosomes and lysosomes, causes a loss of Ser-211 phosphorylation and concomitant nuclear localization of TFEB. We found that while mTORC1 activity toward S6K1, as well as other major mTORC1 substrates, is not impaired, PIKfyve inhibition specifically impedes the interaction of TFEB with mTORC1. This suggests that mTORC1 activity on TFEB is selectively inhibited due to loss of mTORC1 access to TFEB. In addition, we found that TFEB activation during inhibition of PIKfyve relies on the ability of protein phosphatase 2A (PP2A) but not calcineurin/PPP3 to dephosphorylate TFEB Ser-211. Thus when PIKfyve is inhibited, PP2A is dominant over mTORC1 for control of TFEB phosphorylation at Ser-S211. Together these findings suggest that mTORC1 and PP2A have opposing roles on TFEB via phosphorylation and dephosphorylation of Ser-211, respectively, and further that PIKfyve inhibits TFEB activity by facilitating mTORC1-dependent phosphorylation of TFEB.

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References

Ikonomov O, Sbrissa D, Mlak K, Kanzaki M, Pessin J, Shisheva A . Functional dissection of lipid and protein kinase signals of PIKfyve reveals the role of PtdIns 3,5-P2 production for endomembrane integrity. J Biol Chem. 2001; 277(11):9206-11. DOI: 10.1074/jbc.M108750200. View

Raben N, Puertollano R . TFEB and TFE3: Linking Lysosomes to Cellular Adaptation to Stress. Annu Rev Cell Dev Biol. 2016; 32:255-278. PMC: 6490169. DOI: 10.1146/annurev-cellbio-111315-125407. View

Yao Y, Jones E, Inoki K . Lysosomal Regulation of mTORC1 by Amino Acids in Mammalian Cells. Biomolecules. 2017; 7(3). PMC: 5618232. DOI: 10.3390/biom7030051. View

Ebner M, Koch P, Haucke V . Phosphoinositides in the control of lysosome function and homeostasis. Biochem Soc Trans. 2019; 47(4):1173-1185. DOI: 10.1042/BST20190158. View

McCartney A, Zolov S, Kauffman E, Zhang Y, Strunk B, Weisman L . Activity-dependent PI(3,5)P2 synthesis controls AMPA receptor trafficking during synaptic depression. Proc Natl Acad Sci U S A. 2014; 111(45):E4896-905. PMC: 4234577. DOI: 10.1073/pnas.1411117111. View

Ikonomov O, Sbrissa D, Venkatareddy M, Tisdale E, Garg P, Shisheva A . Class III PI 3-kinase is the main source of PtdIns3P substrate and membrane recruitment signal for PIKfyve constitutive function in podocyte endomembrane homeostasis. Biochim Biophys Acta. 2015; 1853(5):1240-50. PMC: 4431544. DOI: 10.1016/j.bbamcr.2015.01.008. View

Lim C, Zoncu R . The lysosome as a command-and-control center for cellular metabolism. J Cell Biol. 2016; 214(6):653-64. PMC: 5021098. DOI: 10.1083/jcb.201607005. View

Zhitomirsky B, Yunaev A, Kreiserman R, Kaplan A, Stark M, Assaraf Y . Lysosomotropic drugs activate TFEB via lysosomal membrane fluidization and consequent inhibition of mTORC1 activity. Cell Death Dis. 2018; 9(12):1191. PMC: 6294013. DOI: 10.1038/s41419-018-1227-0. View

Giridharan S, Luo G, Rivero-Rios P, Steinfeld N, Tronchere H, Singla A . Lipid kinases VPS34 and PIKfyve coordinate a phosphoinositide cascade to regulate retriever-mediated recycling on endosomes. Elife. 2022; 11. PMC: 8816382. DOI: 10.7554/eLife.69709. View

10.

Jeon K, Jono H, Miller C, Cai Y, Lim S, Liu X . Ca2+/calmodulin-stimulated PDE1 regulates the beta-catenin/TCF signaling through PP2A B56 gamma subunit in proliferating vascular smooth muscle cells. FEBS J. 2010; 277(24):5026-39. PMC: 3059601. DOI: 10.1111/j.1742-4658.2010.07908.x. View

11.

Ogunbayo O, Duan J, Xiong J, Wang Q, Feng X, Ma J . mTORC1 controls lysosomal Ca release through the two-pore channel TPC2. Sci Signal. 2018; 11(525). PMC: 6055479. DOI: 10.1126/scisignal.aao5775. View

12.

Cai X, Xu Y, Cheung A, Tomlinson R, Alcazar-Roman A, Murphy L . PIKfyve, a class III PI kinase, is the target of the small molecular IL-12/IL-23 inhibitor apilimod and a player in Toll-like receptor signaling. Chem Biol. 2013; 20(7):912-21. PMC: 4878021. DOI: 10.1016/j.chembiol.2013.05.010. View

13.

Sardiello M, Palmieri M, di Ronza A, Medina D, Valenza M, Gennarino V . A gene network regulating lysosomal biogenesis and function. Science. 2009; 325(5939):473-7. DOI: 10.1126/science.1174447. View

14.

Roczniak-Ferguson A, Petit C, Froehlich F, Qian S, Ky J, Angarola B . The transcription factor TFEB links mTORC1 signaling to transcriptional control of lysosome homeostasis. Sci Signal. 2012; 5(228):ra42. PMC: 3437338. DOI: 10.1126/scisignal.2002790. View

15.

Duex J, Nau J, Kauffman E, Weisman L . Phosphoinositide 5-phosphatase Fig 4p is required for both acute rise and subsequent fall in stress-induced phosphatidylinositol 3,5-bisphosphate levels. Eukaryot Cell. 2006; 5(4):723-31. PMC: 1459661. DOI: 10.1128/EC.5.4.723-731.2006. View

16.

Hasegawa J, Strunk B, Weisman L . PI5P and PI(3,5)P: Minor, but Essential Phosphoinositides. Cell Struct Funct. 2017; 42(1):49-60. PMC: 5846621. DOI: 10.1247/csf.17003. View

17.

Dienstmann R, Rodon J, Serra V, Tabernero J . Picking the point of inhibition: a comparative review of PI3K/AKT/mTOR pathway inhibitors. Mol Cancer Ther. 2014; 13(5):1021-31. DOI: 10.1158/1535-7163.MCT-13-0639. View

18.

Napolitano G, Esposito A, Choi H, Matarese M, Benedetti V, Di Malta C . mTOR-dependent phosphorylation controls TFEB nuclear export. Nat Commun. 2018; 9(1):3312. PMC: 6098152. DOI: 10.1038/s41467-018-05862-6. View

19.

Chen L, Wang K, Long A, Jia L, Zhang Y, Deng H . Fasting-induced hormonal regulation of lysosomal function. Cell Res. 2017; 27(6):748-763. PMC: 5518872. DOI: 10.1038/cr.2017.45. View

20.

Kim E, Goraksha-Hicks P, Li L, Neufeld T, Guan K . Regulation of TORC1 by Rag GTPases in nutrient response. Nat Cell Biol. 2008; 10(8):935-45. PMC: 2711503. DOI: 10.1038/ncb1753. View