Abstract
SKP1-CUL1-F-box (SCF) ubiquitin ligases play fundamental roles in cellular functions. Typically, substrate phosphorylation is required for SCF recognition and subsequent degradation. However, phospho-dependent substrates remain largely unidentified. Here, using quantitative phoshoproteome approach, we performed a system-wide investigation of phospho-dependent SCF substrates. This strategy identified diverse phospho-dependent candidates. Biochemical verification revealed a mechanism by which SCFFBXO22 recognizes the motif XXPpSPXPXX as a conserved phosphodegron to target substrates for destruction. We further demonstrated BAG3, a HSP70 co-chaperone, is a bona fide substrate of SCFFBXO22. FBXO22 mediates BAG3 ubiquitination and degradation that requires ERK-dependent BAG3 phosphorylation at S377. FBXO22 depletion or expression of a stable BAG3 S377A mutant promotes tumor growth via defects in apoptosis and cell cycle progression in vitro and in vivo. In conclusion, our study identified broad phosphorylation-dependent SCF substrates and demonstrated a phosphodegron recognized by FBXO22 and a novel ERK-FBXO22-BAG3 axis involved in tumorigenesis.
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Data availability
All data needed to evaluate the conclusions in the paper are present in the paper and/or the supplementary materials. Mass spectrometry data have been deposited to the iProX (84), a full member of the ProteomeXchange consortium (http://www.iprox.cn), accession ID: IPX0001809000.
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Conceptualization, MT and BL; Methodology, PL, XC, SL, and XJ; Formal Analysis, XC, LZ, DN, and ZL; Investigation, PL, XC, SL, XJ, WD, LZ, XW, YC, JJ, WL, and LP; Resources, MT, BL, MH, and JZ; Data Curation, PL, XC, SL, and XJ; Writing, MT, BL, PL, and XC; Visualization, PL, XC, SL, WD, and XJ; Supervision, MT and BL; Funding Acquisition, MT, BL, WD, and JZ; All authors reviewed and approved the manuscript.
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Our studies did not include human participants, human data, or human tissue. The animal studies were conducted in compliance with the Institutional Animal Care and Use Committee of institutional ethical guidelines in Shanghai Institute of Materia Medica, Chinese Academy of Sciences (2019-03-GMY-10).
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This work was supported by the Natural Science Foundation of China (Nos. 32071432, 81872888, 81821005, 91753203, 81773018, 71473074), National Key R&D Program of China (No. 2020YFE020220), the National Science & Technology Major Project “Key New Drug Creation and Manufacturing Program” (No. 2018ZX09711002-004), the Strategic Priority Research Program of the Chinese Academy of Sciences (No. XDA12050406), the Shanghai Science and Technology Committee (No. 19JC1416300), Innovative Research Team of High-Level Local Universities in Shanghai (SSMU-ZDCX20181202) and the K. C. Wong Education Foundation.
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Liu, P., Cong, X., Liao, S. et al. Global identification of phospho-dependent SCF substrates reveals a FBXO22 phosphodegron and an ERK-FBXO22-BAG3 axis in tumorigenesis. Cell Death Differ 29, 1–13 (2022). https://doi.org/10.1038/s41418-021-00827-7
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DOI: https://doi.org/10.1038/s41418-021-00827-7
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