Parkin ubiquitinates phosphoglycerate dehydrogenase to suppress serine synthesis and tumor progression

作者信息Juan Liu, Cen Zhang, Hao Wu, Xiao-Xin Sun, Yanchen Li, Shan Huang, Xuetian Yue, Shou-En Lu, Zhiyuan Shen, Xiaoyang Su, Eileen White, Bruce G Haffty, Wenwei Hu, Zhaohui Feng
PMID32478681
期刊J Clin Invest
发布时间2020-06-01
DOI10.1172/JCI132876

摘要

Phosphoglycerate dehydrogenase (PHGDH), the first rate-limiting enzyme of serine synthesis, is frequently overexpressed in human cancer. PHGDH overexpression activates serine synthesis to promote cancer progression. Currently, PHGDH regulation in normal cells and cancer is not well understood. Parkin, an E3 ubiquitin ligase involved in Parkinson's disease, is a tumor suppressor. Parkin expression is frequently downregulated in many types of cancer, and its tumor-suppressive mechanism is poorly defined. Here, we show that PHGDH is a substrate for Parkin-mediated ubiquitination and degradation. Parkin interacted with PHGDH and ubiquitinated PHGDH at lysine 330, leading to PHGDH degradation to suppress serine synthesis. Parkin deficiency in cancer cells stabilized PHGDH and activated serine synthesis to promote cell proliferation and tumorigenesis, which was largely abolished by targeting PHGDH with RNA interference, CRISPR/Cas9 KO, or small-molecule PHGDH inhibitors. Furthermore, Parkin expression was inversely correlated with PHGDH expression in human breast cancer and lung cancer. Our results revealed PHGDH ubiquitination by Parkin as a crucial mechanism for PHGDH regulation that contributes to the tumor-suppressive function of Parkin and identified Parkin downregulation as a critical mechanism underlying PHGDH overexpression in cancer.

实验方法

产品清单

名称品牌货号
QuikChange II XL定点诱变试剂盒Agilent Technologies--
PLKO.1潮霉素慢病毒shRNA载体Addgene24150
pSpCas9n(BB)-2A-GFP质粒Addgene48140
抗Flag抗体----
抗Flag磁珠----
抗Myc琼脂糖磁珠Santa Cruz Biotechnology Inc.sc-40
抗Flag琼脂糖磁珠Sigma-AldrichA2220
抗Parkin抗体Cell Signaling Technology4211
抗PHGDH抗体Cell Signaling Technology66350
RIPA裂解缓冲液Thermo Fisher Scientific--
Bradford试剂----
抗Flag-M2抗体Sigma-AldrichF1804
抗GAPDH抗体Santa Cruz Biotechnology Inc.sc-47724
抗HA抗体Sigma-Aldrich11867423001
抗泛素抗体Santa Cruz Biotechnology Inc.sc-8017
抗GST抗体Santa Cruz Biotechnology Inc.sc-138
抗His抗体Santa Cruz Biotechnology Inc.sc-803
抗LC3B抗体Cell Signaling Technology2775
RNeasy试剂盒QIAGEN--
TaqMan逆转录试剂盒Applied Biosystems--
TaqMan PCR混合物Applied Biosystems--
谷胱甘肽-琼脂糖磁珠----
Q Exactive PLUS混合四极杆-轨道阱质谱仪Thermo Fisher Scientific--
UltiMate 3000超高效液相色谱系统----
XBridge BEH酰胺柱Waters--
抗Parkin抗体NovusNB100-91921
抗Ki-67抗体BD Biosciences556003