Cul4A-DDB1–mediated monoubiquitination of phosphoglycerate dehydrogenase promotes colorectal cancer metastasis via increased S-adenosylmethionine
Yajuan Zhang, … , Dawei Li, Weiwei Yang
Yajuan Zhang, … , Dawei Li, Weiwei Yang
Published November 1, 2021
Citation Information: J Clin Invest. 2021;131(21):e146187. https://doi.org/10.1172/JCI146187.
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Research Article Metabolism Oncology

Cul4A-DDB1–mediated monoubiquitination of phosphoglycerate dehydrogenase promotes colorectal cancer metastasis via increased S-adenosylmethionine

Abstract

Although serine metabolism plays a crucial role in the proliferation and survival of tumor cells, how it supports tumor cell migration remains poorly understood. Phosphoglycerate dehydrogenase (PHGDH) catalyzes the oxidation of 3-phosphoglycerate to 3-phosphonooxypyruvate, the first committed step in de novo serine biosynthesis. Here we show that PHGDH was monoubiquitinated by cullin 4A–based E3 ligase complex at lysine 146 in colorectal cancer (CRC) cells, which enhanced PHGDH activity by recruiting a chaperone protein, DnaJ homolog subfamily A member 1, to promote its tetrameric formation, thereby increasing the levels of serine, glycine, and S-adenosylmethionine (SAM). Increased levels of SAM upregulated the expression of cell adhesion genes (laminin subunit gamma 2 and cysteine rich angiogenic inducer 61) by initiating SET domain containing 1A–mediated trimethylation of histone H3K4, thereby promoting tumor cell migration and CRC metastasis. Intriguingly, SAM levels in tumors or blood samples correlated with the metastatic recurrence of patients with CRC. Our finding not only reveals a potentially new role and mechanism of SAM-promoted tumor metastasis but also demonstrates a regulatory mechanism of PHGDH activity by monoubiquitination.

Authors

Yajuan Zhang, Hua Yu, Jie Zhang, Hong Gao, Siyao Wang, Shuxian Li, Ping Wei, Ji Liang, Guanzhen Yu, Xiongjun Wang, Xinxiang Li, Dawei Li, Weiwei Yang

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Figure 3

Cul4A–DDB1–CUL4-associated factor 16 complex mediates the monoubiquitination of PHGDH.

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Cul4A–DDB1–CUL4-associated factor 16 complex mediates the monoubiquitina...
(A) PHGDH proteins were immunoprecipitated from HCT116 cells and the immunoprecipitated complex was further analyzed by liquid chromatography–tandem mass spectrometry for identification of PHGDH-associated proteins. Data represent the means ± SD of 2 independent experiments. (B) HCT116 cells were harvested and PHGDH proteins were immunoprecipitated. (C) HCT116 cells stably expressing SFB-PHGDH WT or K146R were cotransfected with EV or HA-Ub and EV or V5-Cul4A. SFB-PHGDH proteins were pulled down using streptavidin agarose beads. (D) HCT116 cells stably expressing SFB-PHGDH were cotransfected with HA-Ub and siNC, siCul4A, or siDDB1. SFB-PHGDH proteins were pulled down using streptavidin agarose beads. (E) Flag-Cul4A and HA-DCAF16 were immunoprecipitated and purified from HEK293T cells stably expressing Flag-Cul4A or HA-DCAF16. In vitro ubiquitination assay was performed by incubating Flag-Cul4A and HA-DCAF16 with recombinant GST-PHGDH, His-ubiquitin (Ub), His-Uba1 (E1), and His-UbcH6 (E2). (F) HCT116 or SW480 cells stably expressing SFB-PHGDH were transfected with siNC, siCul4A, or siDDB1. SFB-PHGDH proteins were pulled down using streptavidin agarose beads for PHGDH activity measurement. One-way ANOVA with Tukey’s multiple-comparison test. (G) HCT116 cells were transfected with siNC or siCul4A and siDDB1. Cells were harvested for the measurement of intracellular glycine, serine, and SAM (2-tailed Student’s t test). (F and G) Data represent the mean ± SD of 3 biologically independent experiments. (H) IHC staining was performed in paired primary tumors and hepatic metastatic tumors from 10 patients with CRC with anti-Cul4A antibody. Staining scores of Cul4A were compared between primary tumors and metastatic tumors. Left, representative images of IHC staining were shown. Right, semiquantitative scoring was carried out (paired t test, 2 tailed). **P < 0.01. See also Supplemental Figure 3.