Yeast Two Hybrid Screen of a Putative Toxoplasma gondii Cyclin, TGME49_266900

Abstract

In this current research, protein-protein interactions with a putative Toxoplasma gondii cyclin, TGME49_266900 or Cyc6, were discovered via a yeast two hybrid screen. Several putative interacting protein partners were isolated and described from a cDNA library of an asynchronous tachyzoite transcriptome. After false positives were weaned from the study, two proteins were identified as Cyc6 interacting partners. These two proteins are described from the toxodb.org bioinformatic database as a DJ-1 family protein (TGME49_214290) and a ThiF protein (TGME49_314890). Interestingly, the interacting DJ-1 protein has been shown in previous research to play a role in T. gondii microneme secretion. Additionally, ThiF proteins share distinct traits with E1 enzymes at the start of the ubiquitin pathway in eukaryotes. After no evidence of an interacting CDK partner for Cyc6 was obtained, a direct experiment was conducted testing for an interaction between Cyc6 and a putative CDK with expression levels notably higher in bradyzoites instead of tachyzoites. The outcome of this experiment showed no interaction between Cyc6 and the putative CDK. Although no interacting CDK partner was evident from this yeast two hybrid screen, two proteins were found to display a strong and biologically relevant interaction with the putative cyclin of interest. Future studies regarding Cyc6 should explore potential noncanonical roles for this putative cyclin in not only tachyzoites, but also bradyzoites and the purpose of Cyc6\u27s interaction with the two proteins discovered from this screen

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