Elucidating the Polyphosphate Signaling Pathway in Dictyostelium discoideum

Abstract

Polyphosphate acts as a chalone in single celled Dictyostelium discoideum, accumulation of the extracellular signal in the external environment causes cells to halt proliferation and proceed to the next stage of their life cycle. Not much is known about the signaling pathway that occurs within Dictyostelium. The Gomer Lab endeavors to elucidate the polyphosphate signaling pathway. In past semesters, pools of Dictyostelium mutants were generated using Restriction Enzyme-Mediated Integration (REMI) and screened for resistance to poly-p. The genomic DNA of the mutants determined to be resistant was extracted and analyzed. BLAST was used to identify protein matches. The matches selected for verification were irlD and torA. AX2 cells were set as a negative control and gefA- and the AAA-ATPase cells were set as the positive control. Knockout mutants were grown and screened for resistance to poly-p. The results were not statistically significant and more testing is needed. It is most likely that the targets are only slightly resistant to poly-p and have other proteins that contribute to the cell’s sensitivity.