DNA-binding protein PfAP2-P regulates parasite pathogenesis during malaria parasite blood stages
Amit Kumar Subudhi, Judith L. Green, Rohit Satyam, Rahul Salunke, Todd Lenz, Muhammad Shuaib, Ioannis Isaioglou, Steven Abel, Mohit Gupta, Luke Esau, Tobias Mourier, Raushan Nugmanova, Sara Mfarrej, Rupali Shivapurkar, Zenaida Stead, Fathia Ben Rached, Yogesh Ostwal, Rachid Sougrat, Ashraf Dada, Abdullah Fuaad Kadamany, Wolfgang Fischle, Jasmeen S. Merzaban, Ellen Knuepfer, David Ferguson, Ishaan Gupta, Karine G. Le Roch, Anthony A. Holder, Arnab Pain
Abstract
Malaria-associated pathogenesis such as parasite invasion, egress, host cell remodelling and antigenic variation requires concerted action by many proteins, but the molecular regulation is poorly understood. Here we have characterized an essential Plasmodium-specific Apicomplexan AP2 transcription factor in Plasmodium falciparum (PfAP2-P; pathogenesis) during the blood-stage development with two peaks of expression. An inducible knockout of gene function showed that PfAP2-P is essential for trophozoite development, and critical for var gene regulation, merozoite development and parasite egress. Chromatin immunoprecipitation sequencing data collected at timepoints matching the two peaks of pfap2-p expression demonstrate PfAP2-P binding to promoters of genes controlling trophozoite development, host cell remodelling, antigenic variation and pathogenicity. Single-cell RNA sequencing and fluorescence-activated cell sorting revealed de-repression of most var genes in Δpfap2-p parasites. Δpfap2-p parasites also overexpress early gametocyte marker genes, indicating a regulatory role in sexual stage conversion. We conclude that PfAP2-P is an essential upstream transcriptional regulator at two distinct stages of the intra-erythrocytic development cycle.