Characterization Plasmodium yoelii Tudor Staphylococcal Nuclease
Open Access
- Author:
- Smith, Olivia
- Area of Honors:
- Microbiology
- Degree:
- Bachelor of Science
- Document Type:
- Thesis
- Thesis Supervisors:
- Scott Eugene Lindner, Thesis Supervisor
Paul Babitzke, Thesis Honors Advisor - Keywords:
- Malaria
TSN
translational repression
Plasmodium - Abstract:
- Plasmodium parasites are the causative agent of malaria, a disease which afflicted an estimated 219 million people in 2017. The life cycle of these parasites depends on the ability to transmit to mosquitoes, which can then re-infect a host; as such, transmission is a highly regulated process. Through use of Plasmodium yoelii, a strain of the parasite which infects rodents, we are investigating the role of a highly conserved tudor staphylococcal nuclease (TSN) in the transmission process. In other organisms, TSNs are involved in gene expression, but in P. yoelii, the function of PyTSN1 is not well understood. Our lab has previously demonstrated that PyTSN1 interacts with translational repressors ALBA4 and CITH, and the CAF1/CCR4/NOT complex, all of which are involved in transmission of malaria parasites to mosquitoes. To better characterize this protein, I have generated three transgenic parasite lines to identify the function, localization, and protein-protein interactions of PyTSN1 through gene deletion, fluorescent tagging, and proteomics approaches, respectively. Immunofluorescence assays suggest that PyTSN1 is primarily located in the cytoplasm of asexual blood stage parasites and gametocytes. Additionally, PyTSN1 appears to colocalize with CITH, a translational repressor found abundantly in female gametocytes. Protein-protein interactions were determined through the use of a proximity labelling system in which biotin is used as a tag to mark proximal proteins to PyTSN1. Finally, the pytsn1- transgenic parasite line is being analyzed in order to determine if the lack of PyTSN1 conveys any sort of defect in the parasite life cycle. The ultimate goal of this project is to better understand the localization, function, and interactions of PyTSN1 in order to better understand the role this protein plays in transmission of the parasite.