Date of Award

2006

Degree Type

Thesis

Degree Name

Master of Science

Program

Biochemistry

Supervisor

Dr. Fred Dick

Second Advisor

Dr. Eric Ball

Third Advisor

Dr. Gabriel DiMattia

Abstract

The Retinoblastoma tumour suppressor protein (pRB) functions to negatively regulate cell proliferation by inhibiting E2F transcription factor activity. Additionally, an essential function of pRB in the inhibition of apoptosis induced by E2F1 has been documented. The mechanism dictating the ability of pRB to differentially regulate these opposing E2F functions has remained largely elusive. In this thesis I describe the generation of an RB mutant protein that is defective in mediating an E2Fl-t Specific’ binding interaction. This protein retains the ability to inhibit both E2F-mediated transcription and cell cycle entry but is defective in apoptotic regulation, these functions are in contrast to those of the previously described E2F-‘General’ pRB binding mutant. Interestingly, pRB-E2F ‘General’ and ‘Specific’ complexes exhibit different DNA binding specificities. Furthermore, the E2Fl-‘Specific’ domain is uniquely conserved among pRB homologues in higher eukaryotes, highlighting the importance of this apoptotic regulatory domain to pRB function.

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