The Role of the QA Repeat Domain of TCERG1 in the Inhibition of C/EBPα Activity
Transcription elongation regulator 1 (TCERG1) has previously been demonstrated to be an inhibitor of the transactivation and growth arrest activities of CCAAT/Enhancer binding protein alpha (C/EBPα). Furthermore, TCERG1 had been demonstrated to become relocalized from nuclear speckles to the pericentromeric regions where C/EBPα resides when both proteins are co-expressed in the cell. This thesis demonstrates that the deletion of a unique, imperfect series of 38 glutamine-alanine (QA) repeats near the amino terminus of TCERG1 is able to abrogate the ability of TCERG1 to inhibit C/EBPα-mediated growth arrest, the physical interaction between TCERG1 and C/EBPα, and the relocalization of TCERG1 from nuclear speckles when C/EBPα is co-expressed in the cell. The deletion of the QA domain demonstrated that there was a threshold amount of QA repeats required in TCERG1 for the relocalization and growth arrest inhibitory activities between TCERG1 and C/EBPα. It was demonstrated that between 11 and 20 QA repeats were required in TCERG1 to produce the relocalization from nuclear speckles or to be able to inhibit C/EBPα-mediated growth arrest. The physical interaction of TCERG1 and C/EBPα as examined by co-immunoprecipitation was also found to be QA dependent, with a diminishing interaction observed as the number of QA repeats in TCERG1 were reduced. However, experiments examining the isolated QA domain indicated that it was insufficient to relocalize an mCherry fluorescent protein fusion to either the nucleus or to pericentromeric regions where C/EBPα is concentrated. This inability to produce relocalization suggests that the QA domain requires another domain or domains from TCERG1 to mediate the relocalization activity. When expressed with the WT TCERG1, ΔQA TCERG1 was able to act in a dominant negative manner, preventing the relocalization of the WT TCERG1 protein to pericentromeric domains. Interestingly, the transactivation inhibitory activities of TCERG1 on C/EBPα do not appear to require the QA domain, but rather are localized to the carboxy half of TCERG1, somewhere within amino acids 612-1098. The data obtained provides the first report of a role for this unique QA repeat domain.
DegreeMaster of Science (M.Sc.)
SupervisorRoesler, William J.
CommitteeLukong, Kiven E.; Stone, Scot J.; Khandelwal, Ramji; Eskiw, Christopher H.
Copyright DateAugust 2015