Characterization of the essential chromatin remodeling enzyme pfSnf2L in Plasmodium falciparum

The malaria causing parasite Plasmodium falciparum (Pf) is an unicellular eukaryote with a complex life cycle between human host and mosquito vector. Each life cycle stage can be characterized by a specific transcriptional profile, suggesting a complex transcriptional regulation mechanism. The parasite is unique, having the AT-richest genome sequenced to date, the most divergent histone proteins in the eukaryotic domain, resulting in a lack of global nucleosome positioning and exhibiting a 10-fold reduction in number of transcription factors, when compared to organisms of similar complexity. Therefore, it is not understood how the tightly regulated gene expression profile during developmental stage transition is established. It is suggested that the regulation of the gene expression cascade depends on epigenetic mechanisms involving nucleosome remodeling.In contrast to other eukaryotes, Pf has a non-redundant set of chromatin remodeling enzymes. This allows the study of the function of individual classes of chromatin remodelers in the cell by inducible knockout systems and associated functional studies. The ISWI-type enzyme pfSnf2L was chosen, inducible knockout parasite lines were created, and it was shown that pfSnf2L is an essential enzyme for parasite survival. Conditional deletion of pfSnf2L resulted in parasites unable to exit the erythrocyte. In good agreement with this phenotype, gene expression analysis demonstrated that exportome genes are not activated at the correct stage of the intraerythrocytic cycle, in the absence of pfSnf2L. This conditional mutant will allow addressing the effects of pfSnf2L on chromatin structure and gene regulation during the erythrocyte life cycle of Pf. The study is combined with the functional characterization of the recombinant pfSnf2L enzyme, to draw a mechanistic picture of the cellular chromatin changes. The aim is to unravel the mechanism of chromatin remodeling and nucleosome positioning on gene expression networks and phenotypical changes of Plasmodium falciparum.

Project details

Basic Science

Jan 2024

Germany