The ubiquitin E3 ligase ARIH1 regulates hnRNP E1 protein stability, EMT and breast cancer progression

• Published in: Oncogene 2022-03, Vol.41 (12), p.1679-1690
• Main Authors: Howley, Breege V., Mohanty, Bidyut, Dalton, Annamarie, Grelet, Simon, Karam, Joseph, Dincman, Toros, Howe, Philip H.
• Format: Article
• Language: English
• Subjects: 13; 13/106; 13/89; 14/63; 38/111; 38/77; 631/67/1347; 631/67/322; 64/60; 82/51; 82/58; 82/80; Apoptosis; Breast cancer; Breast Neoplasms - genetics; Cell Biology; Cell Line, Tumor; E1 protein; Epithelial cells; Epithelial-Mesenchymal Transition - genetics; Female; Gene expression; Heterogeneous-Nuclear Ribonucleoproteins - genetics; Human Genetics; Humans; Internal Medicine; Mammary gland; Medicine; Medicine & Public Health; Mesenchyme; Metastases; Oncology; Post-transcription; Protein Stability; Proteins; RNA-binding protein; RNA-Binding Proteins - metabolism; Tumors; Ubiquitin; Ubiquitin - metabolism; Ubiquitin-protein ligase; Ubiquitin-Protein Ligases - genetics; Ubiquitin-Protein Ligases - metabolism; Ubiquitination
• ISSN: 0950-9232; 1476-5594
• DOI: 10.1038/s41388-022-02199-9

The epithelial to mesenchymal transition (EMT), a process that is aberrantly activated in cancer and facilitates metastasis to distant organs, requires coordinated transcriptional and post-transcriptional control of gene expression. The tumor-suppressive RNA binding protein, hnRNP-E1, regulates splicing and translation of EMT-associated transcripts and it is thought that it plays a major role in the control of epithelial cell plasticity during cancer progression. We have utilized yeast 2 hybrid screening to identify novel hnRNP-E1 interactors that play a role in regulating hnRNP-E1; this approach led to the identification of the E3 ubiquitin ligase ARIH1. Here, we demonstrate that hnRNP-E1 protein stability is increased upon ARIH1 silencing, whereas, overexpression of ARIH1 leads to a reduction in hnRNP-E1. Reduced ubiquitination of hnRNP-E1 detected in ARIH1 knockdown (KD) cells compared to control suggests a role for ARIH1 in hnRNP-E1 degradation. The identification of hnRNP-E1 as a candidate substrate of ARIH1 led to the characterization of a novel function for this ubiquitin ligase in EMT induction and cancer progression. We demonstrate a delayed induction of EMT and reduced invasion in mammary epithelial cells silenced for ARIH1. Conversely, ARIH1 overexpression promoted EMT induction and invasion. ARIH1 silencing in breast cancer cells significantly attenuated cancer cell stemness in vitro and tumor formation in vivo. Finally, we utilized miniTurboID proximity labeling to identify novel ARIH1 interactors that may contribute to ARIH1’s function in EMT induction and cancer progression.