Toxoplasma gondii is an Apicomplexan parasite present in nearly one third of the world’s population, and is the causative agent of Toxoplasmosis. T. gondii relies on calcium signaling in order to maintain calcium concentrations and execute invasion linked traits (ILTs). These ILTs involve invasion proteins which allow it to establish infection within a host cell. Calcium binding proteins (CBPs) therefore serve as a critical link between calcium regulation and the lytic cycle. This study characterized putative CBPs containing EF-hand domains and featuring high transcript expression. CBP over-expression (OE) mutants exhibited stage-dependent localization patterns at notable sites of calcium-dependent invasion machinery and organelles. CBP1 was found to localize to the membrane of a known calcium store, the plant-like vacuole (PLV). We developed an antibody to CBP1 and saw that it bound to the protein of interest in addition to indeterminate high molecular weight proteins. We obtained CRISPR/Cas9 CBP1 knockout mutants and performed phenotypic assays to observe short-term invasion efficiency, long-term lytic cycle dynamics, and real-time calcium and pH regulation. Overall, we saw that both the over-expression and absence of CBP1 disrupted calcium regulation, pH regulation, and diminished virulence. This study served to illuminate the critical role CBPs occupy in calcium regulation in T. gondii, and their value as novel drug targets in the treatment of parasitic infections.
|Advisor:||Pace, Douglas A.|
|Commitee:||Gharakhanian, Editte, Fraser, Deborah|
|School:||California State University, Long Beach|
|School Location:||United States -- California|
|Source:||MAI 81/3(E), Masters Abstracts International|
|Subjects:||Biology, Molecular biology|
|Keywords:||Calcium binding proteins, Calcium regulation, Lytic cycle, Parasite, Tachyzoite, Toxplasma gondii|
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