Functional characterization of an ApiAP2 transcription factor in sexual development of Plasmodium falciparum

Open Access
- Author:
- Suhendra, Michelle
- Area of Honors:
- Biochemistry and Molecular Biology
- Degree:
- Bachelor of Science
- Document Type:
- Thesis
- Thesis Supervisors:
- Manuel Llinas, Thesis Supervisor
Paul Babitzke, Thesis Honors Advisor - Keywords:
- malaria
gene regulation
transcription
transcription regulation
falciparum
plasmodium
apiap2 - Abstract:
- Malaria is a deadly disease that causes over 500,000 deaths every year; the deadliest form of which is caused by the intracellular eukaryotic parasite Plasmodium falciparum. Regulation of genes essential for parasite survival and development is controlled by the major family of sequence-specific transcription factors known as ApiAP2 proteins. While these transcription factors are required for all life cycle stages in the malaria parasite, only a handful of them have been characterized. One of the 27 members of this family (PF14_0271) has an orthologue in the rodent model species Plasmodium berghei that is important for sexual development and parasite transmission. However, PF14_0271 in P. falciparum has not been characterized and has no known function. This project aims to determine the role of PF14_0271 in the sexual development of P. falciparum, including the protein’s time of expression and DNA-binding specificity. To achieve this, a genetically modified P. falciparum line tagged with GFP was generated. Protein expression levels at the different stages of the parasite life cycle were determined by Western Blot and live fluorescence microscopy and DNA-protein interaction was probed by chromatin immunoprecipitation followed by sequencing (ChIP-seq). The results of these experiments show that PF14_0271 is most highly expressed in the early stage of sexual development and it is localized to the nucleus, where it interacts with at least 100 specific genomic loci. Taken together, these results suggest a heretofore unknown role for PF14_0271 in Plasmodium falciparum sexual development.