FUNCTIONAL CHARACTERIZATION OF E3 UBIQUITIN LIGASE TRIP12 IN MAMMALIAN CELLS

Abstract

TRIP12 is an E3 ubiquitin ligase protein that belongs to the E6-AP C-Terminus (HECT) E3 ubiquitin ligase family. It is a multifunctional protein involved in regulating several mechanisms such as chromatin remodeling, cell cycle progression, DNA damage repair, and more by degrading the respective protein substrates. TGF-β is a family of growth factors involved in a several processes including cell proliferation, cell differentiation, and tissue homeostasis in the intestinal epithelium. Previously, in Dr. Omar Khan’s lab, proteomics data shows that TRIP12 binds to a TGF-β pathway gene, Smad4 (unpublished). We set to understand the functional biology of E3 ubiquitin ligase TRIP12 in TGF-β mediated regulation of cell cycle in established cell line models, validate shRNA Trip12 knockdown efficiency, study the role of Trip12 deletion on the transcriptional activity of TGF-β in NIH3T3 mouse fibroblasts, and establish an immunofluorescence protocol for staining mouse intestinal organoids. In this thesis, TGF-β was found to have no effect on growth inhibition in TRIP12 deleted cells. Additionally, Trip12 deletion was found to enhance TGF- β reporter activity in mouse NIH3T3 cells. A protocol for staining mouse intestinal organoids was established.

Date of Award	2022
Original language	American English
Awarding Institution	HBKU College of Health & Life Sciences