How Does Cyclin B1 Influence Cell Size
Standing
Graduate (Masters)
Type of Proposal
Oral Presentation
Proposal
The tumour suppressor protein, Tuberin is implicated in controlling both growth and division of the cell. Growth is controlled by integrating various nutrient sensing pathways to inhibit protein synthesis and therefore, cell size. Our lab has found that Tuberin controls division by binding to and regulating the localization of the G2/M cyclin, Cyclin B1. Preliminary results show that manipulation of Cyclin B1 and Tuberin results in increased Tuberin protein levels. Therefore, I would like to determine how Cyclin B1 affects the biology of the Tuberin protein. My first aim is to elucidate the residues on Cyclin B1 that mediate Tuberin binding. Cyclin B1 phospho-mutants will be created to determine which residues result in abrogation of Tuberin-Cyclin B1 binding. My second aim is to determine the effect of Cyclin B1 binding on Tuberin. The mutants created in aim one will be used to determine the effect of disrupting Tuberin-Cyclin B1 binding on the protein levels of Tuberin. In addition, the proteasome and lysosome will be inhibited to determine what mechanism Cyclin B1 uses to increase Tuberin protein levels. Lastly, I will also use CRISPR-Cas9 to create a fluorescent Tuberin cell line and use live cell microscopy to monitor Tuberin degradation as it progresses through the cell cycle. How a cell balances growth and division is crucial towards understanding cell size regulation.
Grand Challenges
Healthy Great Lakes
Special Considerations
My submission will not be included in the Grand challenges
How Does Cyclin B1 Influence Cell Size
The tumour suppressor protein, Tuberin is implicated in controlling both growth and division of the cell. Growth is controlled by integrating various nutrient sensing pathways to inhibit protein synthesis and therefore, cell size. Our lab has found that Tuberin controls division by binding to and regulating the localization of the G2/M cyclin, Cyclin B1. Preliminary results show that manipulation of Cyclin B1 and Tuberin results in increased Tuberin protein levels. Therefore, I would like to determine how Cyclin B1 affects the biology of the Tuberin protein. My first aim is to elucidate the residues on Cyclin B1 that mediate Tuberin binding. Cyclin B1 phospho-mutants will be created to determine which residues result in abrogation of Tuberin-Cyclin B1 binding. My second aim is to determine the effect of Cyclin B1 binding on Tuberin. The mutants created in aim one will be used to determine the effect of disrupting Tuberin-Cyclin B1 binding on the protein levels of Tuberin. In addition, the proteasome and lysosome will be inhibited to determine what mechanism Cyclin B1 uses to increase Tuberin protein levels. Lastly, I will also use CRISPR-Cas9 to create a fluorescent Tuberin cell line and use live cell microscopy to monitor Tuberin degradation as it progresses through the cell cycle. How a cell balances growth and division is crucial towards understanding cell size regulation.