Research objectives and content
This proposal outlines a method for the identification of novel factors involved in E2F-1 mediated apoptosis. Elucidation of the E2F-1 apoptotic pathway is necessary for our understanding of the controlling mechanisms which determine the fate of a cell, whether it be proliferation or death. A tightly controlled equilibrium must exist within a cell which governs this process. We are concentrating our efforts specifically on the E2F-1 transcription factor due to its significant role in both proliferative and apoptotic mechanisms. E2F-1 is a major contributor to progression of the cell cycle through the G1/S phase transition and a disturbance in this function is potentially capable of inducing tumorigenesis. Alternatively, the tumor suppresser activity of E2F-1 may be regulated by its capacity to induce cellular apoptosis. To approach this problem we have devised a system that involves screening cDNA libraries by retroviral infection of target cells. The stable target cell lines have been generated by transfection with E2F-1 mutants capable of inducing apoptosis in 100% of the population. It is anticipated that with this system we will identify cDNAs whose translation products are capable of rescuing the apoptotic phenotype induced by deregulated expression of E2F-1. The involvement of these genes in tumor formation/prevention can then be assessed through direct interaction with clinicians in the Pathology Department within the host institute.
Training content (objective, benefit and expected impact)
The training received during the progress of this project will allow me to become familiar with all the methods required for the analysis of cell cycle regulation. It is expected that learning the techniques involved in examining cell cycle control and apoptosis will provide me with an excellent opportunity to initiate independent research within this field following the completion of my work at the host institute.
Links with industry / industrial relevance (22)