The RNF214-TEAD-YAP signaling axis promotes hepatocellular carcinoma progression via TEAD ubiquitylation

RNF214 is an understudied ubiquitin ligase with little knowledge of its biological functions or protein substrates. Here we show that the TEAD transcription factors in the Hippo pathway are substrates of RNF214. RNF214 induces non-proteolytic ubiquitylation at a conserved lysine residue of TEADs, enhances interactions between TEADs and YAP, and promotes transactivation of the downstream genes of the Hippo signaling. Moreover, YAP and TAZ could bind polyubiquitin chains, implying the underlying mechanisms by which RNF214 regulates the Hippo pathway. Furthermore, RNF214 is overexpressed in hepatocellular carcinoma (HCC) and inversely correlates with differentiation status and patient survival. Consistently, RNF214 promotes tumor cell proliferation, migration, and invasion, and HCC tumorigenesis in mice. Collectively, our data reveal RNF214 as a critical component in the Hippo pathway by forming a signaling axis of RNF214-TEAD-YAP and suggest that RNF214 is an oncogene of HCC and could be a potential drug target of HCC therapy.


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provided in the source data.The statistical analysis of the overall survival was through the Human Protein Atlas website (https://www.proteinatlas.org/).The statistical analysis of the progression free survival was through the Kaplan Meier Plotter website (http://kmplot.com/analysis/).The Spearman's rank correlation analysis of the TCGA cohort was through the bioinformatic website (https://www.aclbi.com/static/index.html#/).The mass spectrometry data generated in this study have been deposited in the ProteomeXchange Consortium with the dataset identifier PXD049453.
No significant association with gender was observed in the study.
No significant association with race, ethnicity and other socially relevant groupings was observed in the study.
No significant association with age and gender was observed in the study.
Human HCC tissues and adjacent non-tumor tissues microarray chips were created in Department of Pathology, Zhejiang Provincial People's Hospital (China).
The study was approved by the Medical Ethic Committee of Zhejiang Provincial People's Hospital.
The number of mice used in each experimental group was based on prior experience with liver cancer animal models (PMID:35710796; PMID: 34996827).For in vivo experiments, sample size is of n=8-10.For in vitro experiments, we used sample sizes as listed in the corresponding figure legends or figures.We aimed for a number of at least three independent experiments to allow for Student's t test (two sided).
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Yunqing Qiu & Jianping Jin 2024/3/26 Immunofluorescence were taken by by LSM 710 (Zeiss) confocal microscopy.HE HE or or IHC stained sections were scanned with digital section scanner (KF-FL-020).qRT-PCR analysis was performed by by the SYBR green method (YEASEN) on on Bio-Rad CFX96 system.The dual luciferase assay was performed using Dual Luciferase Reporter Assay System (Vazyme) and detected by by the Spark Multimode Microplate readers (TECAN).The Mass Spec were finished on on timsTOF Pro (Bruker), and the data were analyzed by by PEAKS online.The KEGG pathway enrichment analysis was performed using 'clusterProfiler' R package.Image J (version 1.52r) for image analysis, GraphPad Prism (version7) for statistical analysis All data are available in in the main text or or supplementary informations in in the Source Data.Human data derived from the TCGA, Fanjia, and Tiger datasets were nature portfolio | reporting summary April 2023