IndraLab

"It was observed that GST-full-length (FL) Trabid and Trabid (1–354) strongly interacted with Twist1, whereas GST-Trabid (355–708) showed no interaction (Fig.  xref ), indicating that the N-terminal of Trabid should contain a primary interaction domain for Twist1."

"Mechanistically, Trabid forms a complex with Twist1 and specifically cleaves RNF8-induced K63-linked poly-ubiquitin chains from Twist1, which enhances the association of Twist1 with β-TrCP1 and allows for subsequent K48-linked ubiquitination of Twist1."

"Chromatin immunoprecipitation (ChIP) further confirmed that Twist1 directly binds to the Trabid promoter in HCC cells (Fig.  xref ) and clinical HCC tissues (Fig.  xref )."

"It suggests that Trabid-Twist1 inverse regulation in HCC was associated with reduced survival, and that Trabid downregulation in combination with RNF8 or Twist1 overexpression is a stronger predictor of poor survival than Trabid alone (Fig.  xref )."

"Thus, we concluded that Trabid directly interacts with Twist1."

"To elucidate the mechanisms by which Trabid suppresses Twist1, we firstly examined the interaction between Trabid and Twist1."

"We demonstrate the following: (1) Trabid expression is significantly downregulated in HCC cell lines and two independent cohorts of HCC patients’ tissues; (2) Trabid expression is negatively correlated with pathological grading, recurrence, and poor survival; (3) Reintroduction of Trabid in HCC cells significantly decreases HCC growth and pulmonary metastasis; (4) The inhibitory effect of Trabid on HCC development occurs at least partially through regulation of Twist1 activity; (5) Trabid forms a complex with Twist1 and specifically cleaves RNF8-induced K63-linked poly-ubiquitin chains from Twist1, which enhances the association of Twist1 with β-TrCP1 and allows for subsequent K48-linked ubiquitination of Twist1; (6) AKT-mediated phosphorylation at Ser78/Thr117 is essential for the role of Trabid in negatively regulating Twist1 signaling; and (7) Twist1 negatively regulates the promoter activity of Trabid, indicating a double-negative feedback loop."