TY - JOUR
T1 - A tetracycline-repressible transactivator system to study essential genes in malaria parasites
AU - Pino, Paco
AU - Sebastian, Sarah
AU - Kim, Eunbin Arin
AU - Bush, Erin
AU - Brochet, Mathieu
AU - Volkmann, Katrin
AU - Kozlowski, Elyse
AU - Llinás, Manuel
AU - Billker, Oliver
AU - Soldati-Favre, Dominique
N1 - Funding Information:
We are grateful to Thierry Soldati and William Kelley for critical reading of the manuscript, Valerie Polonais and Arnault Graindorge for their technical assistance, and Beth Krizek for providing reagents for the yeast one-hybrid studies. P.P. was supported by the Wellcome Trust, the HHMI, a Gertrude Von Meissner award, and an EVIMalaR fellowship. A.K. was supported by the HHMI. This work is part of the activities of the BioMalPar and EVIMalaR European Networks of Excellence (LSHP-CT-2004-503578 and number 242095) and was supported by the Swiss National Foundation (D.S.), the Medical Research Council (grant number G0501670 to O.B.), the Wellcome Trust (WT098051 to O.B. and WT077502MA to D.S. and O.B., when both were affiliated to the Department of Biological Sciences at Imperial College London), a Marie Curie Fellowship (PIEF-GA-2008-220180 to S.S.), and an EMBO Long Term Fellowship (to M.B.). M.L. is funded by R01 AI076276 and the Arnold and Mabel Beckman Foundation with support from the Centre for Quantitative Biology (P50GM071508).
PY - 2012/12/13
Y1 - 2012/12/13
N2 - 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.
AB - 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.
UR - https://www.scopus.com/pages/publications/84871020221
UR - https://www.scopus.com/pages/publications/84871020221#tab=citedBy
U2 - 10.1016/j.chom.2012.10.016
DO - 10.1016/j.chom.2012.10.016
M3 - Article
C2 - 23245327
AN - SCOPUS:84871020221
SN - 1931-3128
VL - 12
SP - 824
EP - 834
JO - Cell Host and Microbe
JF - Cell Host and Microbe
IS - 6
ER -