Verification and Characterization of Copine D Mutants in Dictyostelium
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To develop more specific and effective cancer therapeutics, scientists are studying copines. Copines are a novel class of calcium-dependent phospholipid-binding proteins called copines. Copines have been implicated in several different types of human cancers, including leukemia, lung cancer, and breast cancer. The Damer lab at Central Michigan University is using the ameboid protozoan Dictyostelium discoideum to study copines. Dictyostelium has six different copine genes, cpnA to cpnF, and is a good model organism for studying fundamental cellular processes. The Damer lab is focused on determining the function of all six copines; most of the research has been focused on CpnA. My research focused on CpnD. My research aimed to 1) verify our mutant and control cpnD cell lines and 2) characterize the mutant phenotype. We were able to verify our cell lines with PCR and begin the characterization of the phenotype of the cpnD mutants using different growth and development assays and observing the Dictyostelium. Our results indicate that CpnD plays a role in cytokinesis, cell adhesion, and development of Dictyostelium. When compared to cpnA- cells, the cpnD mutant cells had opposite phenotypes. cpnD mutant cells have decreased adhesion and precocious development, while cpnA- cells have increased adhesion and delays in development. After characterizing the mutant phenotype for all six copine genes, we will have a better understanding of the roles copines play in cellular functions. This will improve theories about how copines interact with cancer.