Malaria parasites must respond to stresses and environmental signals to perpetuate efficiently during their multistage development in diverse environments. To gain insights into the parasite's stress response mechanisms, we investigated a conserved Plasmodium protein, which we have named plasmoDJ1 on the basis of the presence of a putative cysteine protease motif of the DJ-1/PfpI superfamily, for its activities, potential to respond to stresses and role in parasite development. PlasmoDJ1 is expressed in all intraerythrocytic stages and ookinetes. Its expression was increased 7–9-fold upon heat shock and oxidative stress due to H2O2 and artemisinin; its expression in a stress-sensitive Escherichia coli mutant conferred tolerance against oxidative stress, indicating that plasmoDJ1 has the potential to sense and/or protect from stresses. Recombinant plasmoDJ1 efficiently neutralized H2O2, facilitated renaturation of denatured citrate synthase and showed protease activity, indicating that plasmoDJ1 is a multi-activity protein. Mutation of the catalytic cysteine residue, but not other residues, reduced H2O2-neutralization activity by ~90% and significantly decreased chaperone and protease activities, indicating that these activities are intrinsic to plasmoDJ1. The plasmoDJ1 gene knockout in Plasmodium berghei ANKA attenuated virulence and reduced oocyst production, suggesting a major role for plasmoDJ1 in parasite development, which probably depends on its multiple activities.
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Research Article|
June 26 2014
Genetic ablation of plasmoDJ1, a multi-activity enzyme, attenuates parasite virulence and reduces oocyst production
Neha Singhal;
Neha Singhal
*CSIR-Centre for Cellular and Molecular Biology, Hyderabad-500007, AP, India
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Atul;
Atul
*CSIR-Centre for Cellular and Molecular Biology, Hyderabad-500007, AP, India
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Babu S. Mastan;
Babu S. Mastan
†University of Hyderabad, Hyderabad-500046, AP, India
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Kota Arun Kumar;
Kota Arun Kumar
†University of Hyderabad, Hyderabad-500046, AP, India
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Puran Singh Sijwali
Puran Singh Sijwali
1
*CSIR-Centre for Cellular and Molecular Biology, Hyderabad-500007, AP, India
1To whom correspondence should be addressed (email psijwali@ccmb.res.in).
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Publisher: Portland Press Ltd
Received:
January 17 2014
Revision Received:
April 17 2014
Accepted:
April 23 2014
Accepted Manuscript online:
April 23 2014
Online ISSN: 1470-8728
Print ISSN: 0264-6021
© The Authors Journal compilation © 2014 Biochemical Society
2014
Biochem J (2014) 461 (2): 189–203.
Article history
Received:
January 17 2014
Revision Received:
April 17 2014
Accepted:
April 23 2014
Accepted Manuscript online:
April 23 2014
Citation
Neha Singhal, Atul, Babu S. Mastan, Kota Arun Kumar, Puran Singh Sijwali; Genetic ablation of plasmoDJ1, a multi-activity enzyme, attenuates parasite virulence and reduces oocyst production. Biochem J 15 July 2014; 461 (2): 189–203. doi: https://doi.org/10.1042/BJ20140051
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