title: A tetracycline-repressible transactivator system to study essential genes in malaria parasites.
abstract: A major obstacle in analyzing gene function in apicomplexan parasites is the absence of a practical regulatable expression system. Here, we identified functional transcriptional activation domains within Apicomplexan AP2 (ApiAP2) family transcription factors. These ApiAP2 transactivation domains were validated in blood-, liver-, and mosquito-stage parasites and used to create a robust conditional expression system for stage-specific, tetracycline-dependent gene regulation in Toxoplasma gondii, Plasmodium berghei, and Plasmodium falciparum. To demonstrate the utility of this system, we created conditional knockdowns of two essential P. berghei genes: profilin (PRF), a protein implicated in parasite invasion, and N-myristoyltransferase (NMT), which catalyzes protein acylation. Tetracycline-induced repression of PRF and NMT expression resulted in a dramatic reduction in parasite viability. This efficient regulatable system will allow for the functional characterization of essential proteins that are found in these important parasites.
authors.affiliation: Department of Microbiology and Molecular Medicine, CMU, University of Geneva, 1 Rue Michel-Servet, 1211 Geneva 4, Switzerland.
authors.forename: Paco
authors.initials: P
authors.lastname: Pino
authors.forename: Sarah
authors.initials: S
authors.lastname: Sebastian
authors.forename: Eunbin Arin
authors.initials: EA
authors.lastname: Kim
authors.forename: Erin
authors.initials: E
authors.lastname: Bush
authors.forename: Mathieu
authors.initials: M
authors.lastname: Brochet
authors.forename: Katrin
authors.initials: K
authors.lastname: Volkmann
authors.forename: Elyse
authors.initials: E
authors.lastname: Kozlowski
authors.forename: Manuel
authors.initials: M
authors.lastname: Llinás
authors.forename: Oliver
authors.initials: O
authors.lastname: Billker
authors.forename: Dominique
authors.initials: D
authors.lastname: Soldati-Favre
mesh_terms: D005786:Gene Expression Regulation; D020043:Genes, Essential; D017125:Genes, Protozoan; D005827:Genetics, Microbial / Q000379:methods*; D008967:Molecular Biology / Q000379:methods*; D010962:Plasmodium berghei / Q000235:genetics*; D010963:Plasmodium falciparum / Q000235:genetics*; D013752:Tetracycline / Q000378:metabolism; D014122:Toxoplasma / Q000235:genetics*; D015534:Trans-Activators / Q000096:biosynthesis
chemical_list: D015534:Trans-Activators; D013752:Tetracycline