The acidocalcisome Ca2+-ATPase (TgA1) of Toxoplasma gondii is required for polyphosphate storage, intracellular calcium homeostasis and virulence.
A large proportion of intracellular Ca2+ in Toxoplasma gondii tachyzoites is stored within acidocalcisomes. These organelles are characterized by their acidic nature and high calcium and polyphosphate (polyP) content. The activity of a Ca2+/H+-ATPase named TgA1 may be important for the accumulation of Ca2+ in these organelles. This enzyme belongs to a group of plasma membrane Ca2+-ATPase (PMCA) that lack a calmodulin-binding domain and have vacuolar localization. To investigate the role of this enzyme, we have generated T. gondii mutants deficient in TgA1 through gene disruption. Proliferation of these mutants decreased dramatically because of deficient cell invasion. In addition, these cells had reduced virulence in a mouse model. Biochemical analysis revealed that the cell polyP content was drastically reduced, and the basal calcium levels were increased and unstable. Microneme secretion under the conditions of stimulation by ionophores was altered. Complementation of null mutants with TgA1 restored most functions. In summary, these results establish a link between TgA1, calcium homeostasis, polyP storage and virulence.
Luo
Shuhong
S
Laboratory of Molecular Parasitology, Department of Pathobiology and Center for Zoonoses Research, College of Veterinary Medicine, University of Illinois at Urbana-Champaign, 2001 South Lincoln Avenue, Urbana, IL 61802, USA.
Ruiz
Felix A
FA
Moreno
Silvia N J
SN
D000818:Animals; D002118:Calcium / Q000378:metabolism*; D000252:Calcium-Transporting ATPases / Q000235:genetics / Q000378:metabolism*; D017931:DNA Primers; D016054:DNA, Protozoan / Q000235:genetics; D005816:Genetic Complementation Test; D006706:Homeostasis; D007700:Kinetics; D016366:Open Reading Frames; D016133:Polymerase Chain Reaction; D011122:Polyphosphates / Q000378:metabolism; D015800:Protozoan Proteins / Q000235:genetics / Q000378:metabolism; D011995:Recombination, Genetic; D014122:Toxoplasma / Q000201:enzymology* / Q000472:pathogenicity*; D014774:Virulence
D017931:DNA Primers; D016054:DNA, Protozoan; D011122:Polyphosphates; D015800:Protozoan Proteins; D000252:Calcium-Transporting ATPases; D002118:Calcium