TgVTC2 is involved in polyphosphate accumulation in Toxoplasma gondii

被引:19
作者
Rooney, Peggy J. [1 ]
Ayong, Lawrence [2 ,3 ]
Tobin, Crystal M. [1 ]
Moreno, Silvia N. J. [2 ,3 ]
Knoll, Laura J. [1 ]
机构
[1] Univ Wisconsin, Sch Med & Publ Hlth, Dept Med Microbiol & Immunol, Madison, WI 53706 USA
[2] Univ Georgia, Ctr Trop & Emerging Global Dis, Athens, GA 30602 USA
[3] Univ Georgia, Dept Cellular Biol, Paul D Coverdell Ctr S350A, Athens, GA 30602 USA
关键词
Toxoplasma; Polyphosphate; SPX; VTC; Q-PCR; FIKK kinase; PLASMODIUM-FALCIPARUM; VTC PROTEINS; V-ATPASE; VIRULENCE; ACIDOCALCISOMES; METABOLISM; PYROPHOSPHATASE; TRAFFICKING; REVEALS; VACUOLE;
D O I
10.1016/j.molbiopara.2010.12.012
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Polyphosphate is found in every cell, having roles in diverse processes, including differentiation and response to stress. In this study, we characterize a Toxoplasma gondii mutant containing an insertion within the carboxy-terminal end of a homolog of Saccharomyces cerevisiae Vtc2p, a component of the polyphosphate synthetic machinery. Locus TgVTC2 encodes a 140 kDa protein containing conserved SPX. VTC and transmembrane domains. TgVTC2 localizes in punctate spots within the cytoplasm that do not co-localize with known markers. The TgVTC2 mutant showed dramatically reduced polyphosphate accumulation, a defect restored by introduction of TgVTC2 to the mutant. Insertion within TgVTC2 resulted in increased transcript levels for two loci, including a putative FIKK kinase. These transcript levels were restored to wild-type levels upon complementation with the TgVTC2 locus. The TgYTC2 locus was refractory to knockout, and may be essential. Analysis of this TgVTC2 mutant will facilitate dissection of the T. gondii polyphosphate synthesis pathway. (C) 2011 Elsevier B.V. All rights reserved.
引用
收藏
页码:121 / 126
页数:6
相关论文
共 21 条
[1]   A MALACHITE GREEN PROCEDURE FOR ORTHO-PHOSPHATE DETERMINATION AND ITS USE IN ALKALINE PHOSPHATASE-BASED ENZYME-IMMUNOASSAY [J].
BAYKOV, AA ;
EVTUSHENKO, OA ;
AVAEVA, SM .
ANALYTICAL BIOCHEMISTRY, 1988, 171 (02) :266-270
[2]   The vtc4 gene influences polyphosphate storage, morphogenesis, and virulence in the maize pathogen Ustilago maydis [J].
Boyce, Kylie J. ;
Kretschmer, Matthias ;
Kronstad, James W. .
EUKARYOTIC CELL, 2006, 5 (08) :1399-1409
[3]   A novel family of yeast chaperons involved in the distribution of V-ATPase and other membrane proteins [J].
Cohen, A ;
Perzov, N ;
Nelson, H ;
Nelson, N .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1999, 274 (38) :26885-26893
[4]   A soluble pyrophosphatase, a key enzyme for polyphosphate metabolism in Leishmania [J].
Espiau, B ;
Lemercier, G ;
Ambit, A ;
Bringaud, F ;
Merlin, G ;
Baltz, T ;
Bakalara, N .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2006, 281 (03) :1516-1523
[5]   Ablation of a small transmembrane protein of Trypanosoma brucei (TbVTC1) involved in the synthesis of polyphosphate alters acidocalcisome biogenesis and function, and leads to a cytokinesis defect [J].
Fang, Jianmin ;
Rohloff, Peter ;
Miranda, Kildare ;
Docampo, Roberto .
BIOCHEMICAL JOURNAL, 2007, 407 :161-170
[6]   Discovery of parasite virulence genes reveals a unique regulator of chromosome condensation 1 ortholog critical for efficient nuclear trafficking [J].
Frankel, Matthew B. ;
Mordue, Dana G. ;
Knoll, Laura J. .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2007, 104 (24) :10181-10186
[7]   Catalytic Core of a Membrane-Associated Eukaryotic Polyphosphate Polymerase [J].
Hothorn, Michael ;
Neumann, Heinz ;
Lenherr, Esther D. ;
Wehner, Mark ;
Rybin, Vladimir ;
Hassa, Paul O. ;
Uttenweiler, Andreas ;
Reinhardt, Monique ;
Schmidt, Andrea ;
Seiler, Jeanette ;
Ladurner, Andreas G. ;
Herrmann, Christian ;
Scheffzek, Klaus ;
Mayer, Andreas .
SCIENCE, 2009, 324 (5926) :513-516
[8]   Polyphosphate and phosphate pump [J].
Kulaev, I ;
Kulakovskaya, T .
ANNUAL REVIEW OF MICROBIOLOGY, 2000, 54 :709-734
[9]   A pyrophosphatase regulating polyphosphate metabolism in acidocalcisomes is essential for Trypanosoma brucei virulence in mice [J].
Lemercier, G ;
Espiau, B ;
Ruiz, FA ;
Vieira, M ;
Luo, SH ;
Baltz, T ;
Docampo, R ;
Bakalara, N .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2004, 279 (05) :3420-3425
[10]   The acidocalcisome Ca2+-ATPase (TgA1) of Toxoplasma gondii is required for polyphosphate storage, intracellular calcium homeostasis and virulence [J].
Luo, SH ;
Ruiz, FA ;
Moreno, SNJ .
MOLECULAR MICROBIOLOGY, 2005, 55 (04) :1034-1045