Molecular cloning, organellar targeting and developmental expression of mitochondrial chaperone HSP60 in Toxoplasma gondii

Toursel, Catherine; Dzierszinski, Florence; Bernigaud, Annie; Mortuaire, Marlène; Tomavo, Stanislas

doi:10.1016/s0166-6851(00)00324-8

Cited by 62 publications

(54 citation statements)

References 32 publications

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“…gondii parasites contain a single mitochondrion, typically forming a lasso-shaped structure surrounding the nucleus (Seeber et al, 1998;Melo et al, 2000;Toursel et al, 2000), but the replication of this organelle has not previously been described. To…”

Section: Resultsmentioning

confidence: 99%

“…(C) Daughter IMC complexes then grow to establish two complete daughter parasites, which are about to emerge, acquiring their plasma membrane from the mother. label the T. gondii mitochondrion, the N-terminal 55 amino acids from mitochondrial matrix heat shock protein 60 (HSP60 L ) (Toursel et al, 2000) were fused to the fluorescent protein DsRed and engineered for stable expression in transgenic parasites. Time-lapse microscopy ( Fig.…”

Section: Resultsmentioning

confidence: 99%

“…(4) Construction methods have previously been described for the basic GFP-expressing vector pdhfrΔCAT-GFP, and the secretion vectors ptubP30 L -GFP and ptubP30 L -YFP Matrajt et al, 2002), as well as constructs that label the ER (ptubP30 L -GFP-HDEL) (Hager et al, 1999), Golgi (ptubGRASP-YFP) (Pelletier et al, 2002), IMC (ptubIMC1-YFP) , micronemes (ptubMIC3-GFP) , rhoptries (ptubROP1-CAT-YFP) (Dzierszinski et al, 2004) and apicoplast [ptubACP L -EGFP (Waller et al, 1998); ptubFNR L -YFP (Striepen et al, 2000;Harb et al, 2004); ptubFNR L -DsRed (He et al, 2001)]. For labeling the mitochondrion, sequences encoding the 55 N-terminal residues of T. gondii heatshock protein 60 (Toursel et al, 2000) were amplified from tachyzoite cDNA using primers 5Ј-atgcagatctaaaATGcttgcccgcgcttca-3Ј and 5Ј-cagtcctagggccgagagtgactccgac-3Ј, and introduced as a BglII-AvrII fragment into ptubIMC1-YFP and ptubFNR L -DsRed, yielding ptubHSP60 L -YFP and ptubHSP60 L -DsRed, respectively.…”

Section: Molecular Methodsmentioning

confidence: 99%

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Organellar dynamics during the cell cycle of Toxoplasma gondii

Nishi

Murray

et al. 2008

Journal of Cell Science

247

341

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results in partitioning of the apicoplast, nucleus, endoplasmic reticulum, and finally the mitochondrion, which enters the developing daughters rapidly, but only very late during the division cycle. The specialized secretory organelles (micronemes and rhoptries) form de novo. This distinctive pattern of replication -in which organellar segregation spans ~75% of the cell cycle, completely encompassing S phase -suggests an unusual mechanism of cell cycle regulation. Supplementary material available online at

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Molecular Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Organellar dynamics during the cell cycle of Toxoplasma gondii

Nishi

Murray

et al. 2008

Journal of Cell Science

247

341

View full text Add to dashboard Cite

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“…For example, T. gondii myosins B and C are encoded by alternatively spliced mRNA and differ only in their C-terminal tails (30). Two cDNAs of T. gondii heat shock protein HSP60 corresponding to a single gene have also been isolated and characterized (31). In addition, two isoforms of the T. gondii hypoxanthine-xanthine-guanine phosphoribosyltransferase encoded by a single genetic locus, differing by only a 49-amino acid insertion, have recently been demonstrated (32).…”

Section: Discussionmentioning

confidence: 99%

The Farnesyl-diphosphate/Geranylgeranyl-diphosphate Synthase of Toxoplasma gondii Is a Bifunctional Enzyme and a Molecular Target of Bisphosphonates

Ling

Miranda

et al. 2007

Journal of Biological Chemistry

115

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Farnesyl-diphosphate synthase (FPPS) catalyzes the synthesis of farnesyl diphosphate, an important precursor of sterols, dolichols, ubiquinones, and prenylated proteins. We report the cloning and characterization of two Toxoplasma gondii farnesyl-diphosphate synthase (TgFPPS) homologs. A single genetic locus produces two transcripts, TgFPPS and TgFPPSi, by alternative splicing. Both isoforms were heterologously expressed in Escherichia coli, but only TgFPPS was active. The protein products predicted from the nucleotide sequences have 646 and 605 amino acids and apparent molecular masses of 69.5 and 64.5 kDa, respectively. Several conserved sequence motifs found in other prenyl-diphosphate synthases are present in both TgFPPSs. TgFPPS was also expressed in the baculovirus system and was biochemically characterized. In contrast to the FPPS of other eukaryotic organisms, TgFPPS is bifunctional, catalyzing the formation of both farnesyl diphosphate and geranylgeranyl diphosphate. TgFPPS localizes to the mitochondria, as determined by the co-localisation of the affinity-purified antibodies against the protein with MitoTracker, and in accord with the presence of an N-terminal mitochondria-targeting signal in the protein. This enzyme is an attractive target for drug development, because the order of inhibition of the enzyme by a number of bisphosphonates is the same as that for inhibition of parasite growth. In summary, we report the first bifunctional farnesyl-diphosphate/geranylgeranyldiphosphate synthase identified in eukaryotes, which, together with previous results, establishes this enzyme as a valid target for the chemotherapy of toxoplasmosis.Toxoplasma gondii is a pathogenic protozoan parasite that infects a wide range of vertebrate hosts, including humans. T. gondii has been recognized as a major opportunistic pathogen of fetuses from recently infected mothers and of immunocompromised patients, i.e. those with AIDS. Toxoplasmic encephalitis is associated with high mortality and morbidity and is one of the most common opportunistic infections of the central nervous system in human immunodeficiency virus-infected patients (1). The chemotherapy for toxoplasmosis is not ideal especially for the AIDS patient because there is relapse of the infection if the treatment is halted because of intolerance of the patient to the side effects. Almost all the drugs in use have toxicity associated with their continued use. In addition, most of the recommended treatments do not have an effect on the slow growing bradyzoite forms.Isoprenoids are the most diverse and abundant compounds occurring in nature. Many types of isoprenoids (e.g. steroids, cholesterol, retinoids, carotenoids, ubiquinones, and prenyl groups bound to proteins) are essential components of the cellular machinery of all organisms because of their roles in a variety of biological processes. Despite their structural and functional variety, all isoprenoids derive from a common precursor, isopentenyl diphosphate (IPP), 2 and its isomer, dimethylallyl diphosphate (...

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“…gondii heat shock protein 60 (TgHSP60) is a member of the HSP60 subfamily (Toursel et al, 2000), which plays an important role in intracellular survival and in the differentiation of T. gondii and also mediates its virulence (Shonhai et al, 2011). In addition, TgHSP60 not only serves as a signal for the activation of antigen presenting cells by inducing the release of cytokines and stimulation of the initial immune response, but also acts as a molecular chaperone in the mitochondria (Ma et al, 2009).…”

Section: Introductionmentioning

confidence: 99%

Characterization of the Toxoplasma gondii hsp60 gene sequences from different hosts and geographical locations

et al. 2014

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ABSTRACT. The intracellular protozoan Toxoplasma gondii is one of the most successful parasites, with the ability to invade all warmblooded animals, including humans. T. gondii heat shock protein 60 (TgHSP60) plays an important role in intracellular survival and in the differentiation of the parasite, and is also recognized as being associated with its virulence. In the present study, we examined sequence variation in the hsp60 coding region among five T. gondii isolates from different hosts and geographical regions, which were compared with the corresponding sequences of strains ME49, 76K, and GT1 available in the ToxoDB databases. The length of the T. gondii hsp60 sequence was 1728 bp for all strains, and the A+T content ranged from 41.96 to 42.13%. The sequence alignment of the 8 T. gondii strains identified 20 variable positions (0-1.44%) and showed 1.16% overall sequence 6907 ©FUNPEC-RP www.funpecrp.com.br Genetics and Molecular Research 13 (3): 6906-6911 (2014) Sequence variation in the T. gondii hsp60 gene variation, suggesting a relatively considerable sequence diversity. Phylogenetic analysis of hsp60 sequences using Bayesian inference and maximum parsimony differentiated the two major clonal lineage types into their respective clusters, and thus separated atypical strains from classical genotypes. The results of the present study suggested that the coding region of the hsp60 gene may represent a novel genetic marker for intraspecies phylogenetic analyses of T. gondii.

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Molecular cloning, organellar targeting and developmental expression of mitochondrial chaperone HSP60 in Toxoplasma gondii

Cited by 62 publications

References 32 publications

Organellar dynamics during the cell cycle of Toxoplasma gondii

Organellar dynamics during the cell cycle of Toxoplasma gondii

The Farnesyl-diphosphate/Geranylgeranyl-diphosphate Synthase of Toxoplasma gondii Is a Bifunctional Enzyme and a Molecular Target of Bisphosphonates

Characterization of the Toxoplasma gondii hsp60 gene sequences from different hosts and geographical locations

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