Differential expression of two plant-like enolases with distinct enzymatic and antigenic properties during stage conversion of the protozoan parasite Toxoplasma gondii

Dzierszinski, Florence; Mortuaire, Marlène; Dendouga, Najoua; Popescu, Octavian; Tomavo, Stanislas

doi:10.1006/jmbi.2001.4730

Cited by 110 publications

(104 citation statements)

References 28 publications

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“…T. gondii RH tachyzoites and HXGPRT-deficient strain (National Institutes of Health AIDS Reference and Reagent Repository, Bethesda, MD) were maintained by serial passage in HFF monolayers in Dulbecco's modified Eagle's medium supplemented with 10% fetal calf serum and 20 g/ml gentamicin solution at 37°C in a humid 5% CO 2 atmosphere. Encysted bradyzoites were purified from the brains of OlaHsd or Swiss mice chronically infected with the cyst-forming strain 76 K as described previously (13).…”

Section: Methodsmentioning

confidence: 99%

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Cathepsin Cs Are Key for the Intracellular Survival of the Protozoan Parasite, Toxoplasma gondii

Que

Engel

Ferguson

et al. 2007

Journal of Biological Chemistry

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Cysteine proteases play key roles in apicomplexan invasion, organellar biogenesis, and intracellular survival. We have now characterized five genes encoding papain family cathepsins from Toxoplasma gondii, including three cathepsin Cs, one cathepsin B, and one cathepsin L. Unlike endopeptidases cathepsin B and L, T. gondii cathepsin Cs are exopeptidases and remove dipeptides from unblocked N-terminal substrates of proteins or peptides. TgCPC1 was the most highly expressed cathepsin mRNA in tachyzoites (by real-time PCR), but three cathepsins, TgCPC1, TgCPC2, and TgCPB, were undetectable in in vivo bradyzoites. The specific cathepsin C inhibitor, Gly-Phe-dimethylketone, selectively inhibited the TgCPCs activity, reducing parasite intracellular growth and proliferation. The targeted disruption of TgCPC1 does not affect the invasion and growth of tachyzoites as TgCPC2 is then up-regulated and may substitute for TgCPC1. TgCPC1 and TgCPC2 localize to constitutive secretory vesicles of tachyzoites, the dense granules. T. gondii cathepsin Cs are required for peptide degradation in the parasitophorous vacuole as the degradation of the marker protein, Escherichia coli ␤-lactamase, secreted into the parasitophorous vacuole of transgenic tachyzoites was completely inhibited by the cathepsin C inhibitor. Cathepsin C inhibitors also limited the in vivo infection of T. gondii in the chick embryo model of toxoplasmosis. Thus, cathepsin Cs are critical to T. gondii growth and differentiation, and their unique specificities could be exploited to develop novel chemotherapeutic agents.

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Section: Methodsmentioning

confidence: 99%

“…Total RNA from in vivo encysted bradyzoites was purified with TRIzol (Invitrogen), treated with DNase, and reversetranscribed as described previously (13). Comparative RT-PCR was performed using equal amounts of tachyzoite and bradyzoite cDNAs based on normalization of the amplified cDNA of T. gondii housekeeping actin gene.…”

Section: Methodsmentioning

confidence: 99%

Cathepsin Cs Are Key for the Intracellular Survival of the Protozoan Parasite, Toxoplasma gondii

Que

Engel

Ferguson

et al. 2007

Journal of Biological Chemistry

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“…Approximately 3%-5% showed the highest similarity outside the phylum Apicomplexa to a plant gene, whereas 5%-10% were most similar to animals (Table 3). Previous studies have also highlighted the fact that many metabolic enzymes in T. gondii are plant-like (Dzierszinski et al 2001). These plant-like genes may have originated from an algal endosymbiont that is still represented by the remnant apicoplast (Kohler et al 1997;Roos et al 1999).…”

Section: Phylogenetic Comparison Of Gene Homologiesmentioning

confidence: 99%

Gene Discovery in the Apicomplexa as Revealed by EST Sequencing and Assembly of a Comparative Gene Database

Li¹,

Brunk²,

Kissinger³

et al. 2003

Genome Res.

133

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Large-scale EST sequencing projects for several important parasites within the phylum Apicomplexa were undertaken for the purpose of gene discovery. Included were several parasites of medical importance (Plasmodium falciparum, Toxoplasma gondii) and others of veterinary importance (Eimeria tenella, Sarcocystis neurona, and Neospora caninum). A total of 55,192 ESTs, deposited into dbEST/GenBank, were included in the analyses. The resulting sequences have been clustered into nonredundant gene assemblies and deposited into a relational database that supports a variety of sequence and text searches. This database has been used to compare the gene assemblies using BLAST similarity comparisons to the public protein databases to identify putative genes. Of these new entries, ∼15%-20% represent putative homologs with a conservative cutoff of p < 10 −9 , thus identifying many conserved genes that are likely to share common functions with other well-studied organisms. Gene assemblies were also used to identify strain polymorphisms, examine stage-specific expression, and identify gene families. An interesting class of genes that are confined to members of this phylum and not shared by plants, animals, or fungi, was identified. These genes likely mediate the novel biological features of members of the Apicomplexa and hence offer great potential for biological investigation and as possible therapeutic targets.

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“…2). Alignement analysis also showed that characteristic residues involved in the catalytic activity of the ENO were conserved [15] in all of the sequences presented in Fig. 2.…”

Section: Multiple Amino Acid Sequence Alignment and Phylogenetic Analmentioning

confidence: 73%

Isolation, Cloning and Sequence Analysis of Enolase Enzyme Encoding Gene from Theileria annulata for Assessment of Important Residues of This Enzyme

Akat¹,

Aktaş²,

Dumanlı³

et al. 2014

Kafkas Univ Vet Fak Derg

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SummaryDrug resistance against one of the important antitheilerial drugs has been reported for the first time in 2010. For the aim of developing new antitheilerial drugs or vaccines, enolase gene was isolated from the genomic DNA of Theileria annulata, cloned for the first time in the literature and analyzed at nucleotide and amino acid levels by using different web based tools. These analyses showed that the gene was consisted of 1365 nucleotides including an intron sequence placed between residues 40-41. Restriction enzyme mapping analysis of the cloned gene showed that, base pair changes in TaENO Elazig strain caused differences on cutting and non-cutting restriction enzymes compared to the Ankara strain. These differences may help the identification of different strains by restriction mapping and it may be possible to determine the geological distribution of T. annulata strains in any region. As the comparison of enolase gene sequences from T. annulata and the muscle enolase isoform of the host Bos taurus was made, four different insertions in T. annulata enolase that do not exist in B. taurus enolase was reported as an important discovery of this study. The modeling studies on T. annulata enolase gene showed that these insertions constituted loops that do not exist in B. taurus enolase, suggesting that these loops could be specific binding sites for enzyme inhibitors. Keywords: Theileria annulata, Enolase, Strain identification, Antitheilerial drugs, Structure based drug design, Protein homology modelingTheileria annulata'nın Enolaz Enzimini Kodlayan Geninin, Önemli Rezidülerinin Değerlendirilmesi Amacıyla İzolasyonu, Klonlanması ve Dizi Analizinin Yapılması ÖzetBu yıl ilk kez önemli bir antitheilerial ilaca karşı direnç geliştiği rapor edilmiştir. Yeni antitheilerial ilaçların veya aşıların geliştirilmesi amacıyla Theileria annulata genomik DNA'sından enolaz geni izole edilmiş, literatürde ilk kez klonlanmış ve web tabanlı araçlar kullanılarak hem nükleotid hem de amino asit seviyesinde analiz edilmiştir. Bu analizler genin 1365 nükleotidden oluştuğunu ve 40-41 rezidüleri arasında bir intron dizisi içerdiğini göstermiştir. Klonlanan genin restriksiyon enzim haritalama analizinde, TaENO Elazığ soyundaki baz çifti değişikliklerinin, Ankara soyu ile karşılaştırıldığında, geni kesen ve kesmeyen enzimlerde farklılıklar yarattığı gözlemlenmiştir. Bu farklılıklar, farklı soyların restriksiyon haritalama ile tanımlanmasına yardımcı olabilir ve T. annulata soylarının herhangi bir bölgedeki coğrafik dağılımının belirlenmesini mümkün kılabilir. T. annulata enolaz geni dizisi ile konak Bos taurus'un kas enolaz izoformu karşılaştırıldığında, T. annulata'da bulunan 4 farklı insersiyonun B. taurus enolazında bulunmadığı belirlenmiş ve bu çalışmanın önemli bir bulgusu olarak rapor edilmiştir. T. annulata enolazının modelleme çalışmaları bu insersiyonların B. taurus'ta bulunmayan halkalar oluşturduğunu göstermiş ve bu halkaların enzim inhibitörleri için spesifik bağlanma bölgeleri olabileceği önerilmiştir.

show abstract

Differential expression of two plant-like enolases with distinct enzymatic and antigenic properties during stage conversion of the protozoan parasite Toxoplasma gondii

Cited by 110 publications

References 28 publications

Cathepsin Cs Are Key for the Intracellular Survival of the Protozoan Parasite, Toxoplasma gondii

Cathepsin Cs Are Key for the Intracellular Survival of the Protozoan Parasite, Toxoplasma gondii

Gene Discovery in the Apicomplexa as Revealed by EST Sequencing and Assembly of a Comparative Gene Database

Isolation, Cloning and Sequence Analysis of Enolase Enzyme Encoding Gene from Theileria annulata for Assessment of Important Residues of This Enzyme

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