The Tumour Suppressor Tuberin Coordinating DNA damage and Mitotic Onset.
Standing
Undergraduate
Type of Proposal
Oral Research Presentation
Faculty
Faculty of Science
Faculty Sponsor
Dr. Lisa Porter
Proposal
The cell cycle is a series of events that involve cell growth and division. It is regulated by Cyclins and CDKs, as well as checkpoints that control its progression. Tuberin (TSC2 gene) is a tumour suppressor protein that controls cell growth and cell proliferation. Tuberin mutations are implicated in several cancers and diseases such as tuberous sclerosis complex (TSC), a multisystem genetic disorder that is characterized by the growth of hamartomas. Previous data from our lab has demonstrated Tuberin’s role in controlling miotic onset by binding to the G2/M cyclin, Cyclin B1 according to nutrient availability. My project explores the role of the tuberin in the G2/M transition during DNA damage repair. DNA damage can result from events such as radiation, X-ray, and reactive oxygen species which activates the G2/M checkpoint to restrict mitotic onset and ensure sufficient time to repair damaged DNA. In our studies, NIH3T3 p53 positive cell line cells are being transfected with TSC2-WT or clinical TSC2 mutants that present low affinity for Cyclin B1. The transfected cells will be treated with etoposide, a topoisomerase II drug that induces double-stranded DNA breaks during the G2/M transition. The changes in cell cycle phases are being analyzed by flow cytometry. Our preliminary results have determined the best concentration of etoposide (10uM) and timing (4 hours) to promote G2/M arrest in our experimental conditions. This project will help expand the role of Tuberin during mitotic onset and clarify mechanisms of proliferative diseases as TSC and cancers.
Availability
March 29 (12-1:30), March 30 (12-3), March 31 (12-1:30), April 1 (12:30-3)
The Tumour Suppressor Tuberin Coordinating DNA damage and Mitotic Onset.
The cell cycle is a series of events that involve cell growth and division. It is regulated by Cyclins and CDKs, as well as checkpoints that control its progression. Tuberin (TSC2 gene) is a tumour suppressor protein that controls cell growth and cell proliferation. Tuberin mutations are implicated in several cancers and diseases such as tuberous sclerosis complex (TSC), a multisystem genetic disorder that is characterized by the growth of hamartomas. Previous data from our lab has demonstrated Tuberin’s role in controlling miotic onset by binding to the G2/M cyclin, Cyclin B1 according to nutrient availability. My project explores the role of the tuberin in the G2/M transition during DNA damage repair. DNA damage can result from events such as radiation, X-ray, and reactive oxygen species which activates the G2/M checkpoint to restrict mitotic onset and ensure sufficient time to repair damaged DNA. In our studies, NIH3T3 p53 positive cell line cells are being transfected with TSC2-WT or clinical TSC2 mutants that present low affinity for Cyclin B1. The transfected cells will be treated with etoposide, a topoisomerase II drug that induces double-stranded DNA breaks during the G2/M transition. The changes in cell cycle phases are being analyzed by flow cytometry. Our preliminary results have determined the best concentration of etoposide (10uM) and timing (4 hours) to promote G2/M arrest in our experimental conditions. This project will help expand the role of Tuberin during mitotic onset and clarify mechanisms of proliferative diseases as TSC and cancers.