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Originally published In Press as doi:10.1074/jbc.M102402200 on May 22, 2001

J. Biol. Chem., Vol. 276, Issue 28, 26114-26121, July 13, 2001
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Rapid Changes in Polyphosphate Content within Acidocalcisomes in Response to Cell Growth, Differentiation, and Environmental Stress in Trypanosoma cruzi*

Felix A. Ruiz, Claudia O. Rodrigues, and Roberto DocampoDagger

From the Laboratory of Molecular Parasitology, Department of Pathobiology, University of Illinois at Urbana-Champaign, Urbana, Illinois 61802

Inorganic polyphosphate (polyP) has been identified and measured in different stages of Trypanosoma cruzi. Millimolar levels (in terms of Pi residues) in chains of less than 50 residues long, and micromolar levels in chains of about 700-800 residues long, were found in different stages of T. cruzi. Analysis of purified T. cruzi acidocalcisomes indicated that polyPs were preferentially located in these organelles. This was confirmed by visualization of polyPs in the acidocalcisomes using 4',6-diamidino-2-phenylindole. A rapid increase (within 2-4 h) in the levels of short and long chain polyPs was detected during trypomastigote to amastigote differentiation and during the lag phase of growth of epimastigotes (within 12-24 h). Levels rapidly decreased after the epimastigotes resumed growth. Short and long chain polyP levels rapidly decreased upon exposure of epimastigotes to hypo-osmotic or alkaline stresses, whereas levels increased after hyperosmotic stress. Ca2+ release from acidocalcisomes by a combination of ionophores (ionomycin and nigericin) was associated with the hydrolysis of short and long chain polyPs. In agreement with these results, acidocalcisomes were shown to contain polyphosphate kinase and exopolyphosphatase activities. Together, these results suggest a critical role for these organelles in the adaptation of the parasite to environmental changes.


* This work was supported by National Institutes of Health Grant AI-23259 (to R. D.).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Dagger To whom correspondence should be addressed: Laboratory of Molecular Parasitology, Dept. of Pathobiology, College of Veterinary Medicine, University of Illinois at Urbana-Champaign, 2001 S. Lincoln Ave., Urbana, IL 61802. Tel.: 217-333-3845; Fax: 217-244-7421; E-mail: rodoc@uiuc.edu.


Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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