Evaluation of the cell cycle regulation in Tuberin knockout cell lines
Standing
Undergraduate
Type of Proposal
Oral Research Presentation
Faculty
Faculty of Science
Faculty Sponsor
Lisa A. Porter
Proposal
Tuberin (TSC2) and its binding partner hamartin (TSC1) are tumour suppressors proteins that regulate cell growth and cell proliferation through the mTOR pathway and cell cycle, respectively. Mutations in these genes are related to many proliferative diseases such as Tuberous Sclerosis Complex (TSC) and cancers. Our lab has determined a novel role for TSC2 as a cell cycle regulator at the G2/M transition. TSC2 is able to bind to the G2/M cyclin, Cyclin B1 (CycB1), and retain this cyclin in the cytoplasm, delaying mitotic onset. My project focuses on further understanding the TSC2/CycB1 complex formation and the consequences of this interaction during progression of the cell cycle. I hypothesize that cells lacking TSC2 will have an improper G2/M transition causing morphological and proliferative abnormalities. To test my hypothesis I’m using TSC1 or/and TSC2 knockout HEK293 cells lines acquired from the TSC Biosample Repository to measure the rate of the G2/M transition by flow cytometry, cell proliferation through BrDU assay and cell morphology by immunofluorescence. Preliminary data shows that the HEK293 knockout cells lines present an altered cell cycle prolife when compared to the WT cells. The knockout cell lines present a large population of cells in G1 phase when compared to the WT cell line. To further understand the TSC2 knockout consequences, the cells are being synchronized using thymidine block (S phase blocker) and the cell cycle transition will be monitored for each of the cell cycle phases following release. My project will shed light on the role of TSC2 as a cell cycle regulator and clarify its role as a tumour suppressor protein.
Availability
March 29: 1pm-3pm, March 30: 12pm-1pm, March 31: 1pm-3pm, April 1: 12pm-1pm
Special Considerations
Jose Antonio Roye-Azar is the presenter
Evaluation of the cell cycle regulation in Tuberin knockout cell lines
Tuberin (TSC2) and its binding partner hamartin (TSC1) are tumour suppressors proteins that regulate cell growth and cell proliferation through the mTOR pathway and cell cycle, respectively. Mutations in these genes are related to many proliferative diseases such as Tuberous Sclerosis Complex (TSC) and cancers. Our lab has determined a novel role for TSC2 as a cell cycle regulator at the G2/M transition. TSC2 is able to bind to the G2/M cyclin, Cyclin B1 (CycB1), and retain this cyclin in the cytoplasm, delaying mitotic onset. My project focuses on further understanding the TSC2/CycB1 complex formation and the consequences of this interaction during progression of the cell cycle. I hypothesize that cells lacking TSC2 will have an improper G2/M transition causing morphological and proliferative abnormalities. To test my hypothesis I’m using TSC1 or/and TSC2 knockout HEK293 cells lines acquired from the TSC Biosample Repository to measure the rate of the G2/M transition by flow cytometry, cell proliferation through BrDU assay and cell morphology by immunofluorescence. Preliminary data shows that the HEK293 knockout cells lines present an altered cell cycle prolife when compared to the WT cells. The knockout cell lines present a large population of cells in G1 phase when compared to the WT cell line. To further understand the TSC2 knockout consequences, the cells are being synchronized using thymidine block (S phase blocker) and the cell cycle transition will be monitored for each of the cell cycle phases following release. My project will shed light on the role of TSC2 as a cell cycle regulator and clarify its role as a tumour suppressor protein.