Is cancer an aggregation disease? Generation of p53 mutations.
Authors:Genesee Martinez, Akeema Onalaja
Mentor:Luiza Nogaj, PhD, Associate Professor, Mount St. Mary's College
p53, often called guardian of the genome, is a tumor suppressor made up of four subunits with very specific DNA binding sequences. It was found that p53 is mutated in 50% of all cancers, including lung, breast and prostate cancers. When there is a mutation within this gene, or its binding sequences, the subunits do not function properly and potentially clump together to form aggregates of mutated p53. These aggregates then go on to alter the expression and regulation of other genes, and thus eliminating the cancer fighting ability of this sequence specific protein. The objective of this study is to generate the most common p53 mutations and analyze their ability to bind to DNA. p53 mutants will also be examined for stability and ability to aggregate. To address these goals, we identified and examined the most common p53 mutations in frequently occurring cancers. First we cloned wild-type p53, and then, using QuickChange® Site-Directed Mutagenesis we generated the following mutations: Arg175, Arg248, and Arg273, Gly245, Arg249, and Arg158. Future studies will aim at defining p53 mutant stability and their ability to aggregate.