Structure and expression of three gp82 gene subfamilies of Trypanosoma cruzi

Songthamwat, Dujdow; Kajihara, Kazuo; Kikuchi, Mihoko; Uemura, Hirotsugu; Tran, Sieu Phu Manh; Yanagi, Tetsuo; Higo, Hiroo; Hirayama, Kenji

doi:10.1016/j.parint.2007.05.004

Cited by 6 publications

(4 citation statements)

References 38 publications

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“…Given this, it is important that further parasite genetic characterization and association studies with clinical pathology are carried out. Several important genes that may be involved in the invasion, proliferation and immune evasion of these parasites have been identified [46], [47], and these would be obvious candidate genes for pathogenicity association studies.…”

Section: Resultsmentioning

confidence: 99%

Lineage Analysis of Circulating Trypanosoma cruzi Parasites and Their Association with Clinical Forms of Chagas Disease in Bolivia

Puerto

Nishizawa²,

Kikuchi

et al. 2010

PLoS Negl Trop Dis

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BackgroundThe causative agent of Chagas disease, Trypanosoma cruzi, is divided into 6 Discrete Typing Units (DTU): Tc I, IIa, IIb, IIc, IId and IIe. In order to assess the relative pathogenicities of different DTUs, blood samples from three different clinical groups of chronic Chagas disease patients (indeterminate, cardiac, megacolon) from Bolivia were analyzed for their circulating parasites lineages using minicircle kinetoplast DNA polymorphism.Methods and FindingsBetween 2000 and 2007, patients sent to the Centro Nacional de Enfermedades Tropicales for diagnosis of Chagas from clinics and hospitals in Santa Cruz, Bolivia, were assessed by serology, cardiology and gastro-intestinal examinations. Additionally, patients who underwent colonectomies due to Chagasic magacolon at the Hospital Universitario Japonés were also included. A total of 306 chronic Chagas patients were defined by their clinical types (81 with cardiopathy, 150 without cardiopathy, 100 with megacolon, 144 without megacolon, 164 with cardiopathy or megacolon, 73 indeterminate and 17 cases with both cardiopathy and megacolon). DNA was extracted from 10 ml of peripheral venous blood for PCR analysis. The kinetoplast minicircle DNA (kDNA) was amplified from 196 out of 306 samples (64.1%), of which 104 (53.3%) were Tc IId, 4 (2.0%) Tc I, 7 (3.6%) Tc IIb, 1 (0.5%) Tc IIe, 26 (13.3%) Tc I/IId, 1 (0.5%) Tc I/IIb/IId, 2 (1.0%) Tc IIb/d and 51 (25.9%) were unidentified. Of the 133 Tc IId samples, three different kDNA hypervariable region patterns were detected; Mn (49.6%), TPK like (48.9%) and Bug-like (1.5%). There was no significant association between Tc types and clinical manifestations of disease.ConclusionsNone of the identified lineages or sublineages was significantly associated with any particular clinical manifestations in the chronic Chagas patients in Bolivia.

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Section: Resultsmentioning

confidence: 99%

Lineage Analysis of Circulating Trypanosoma cruzi Parasites and Their Association with Clinical Forms of Chagas Disease in Bolivia

Puerto

Nishizawa²,

Kikuchi

et al. 2010

PLoS Negl Trop Dis

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“…Gp82 expression has been shown to be greatly reduced in the non-virulent T. cruzi strain, CL-14, thereby supporting the hypothesis that this protein is required for cell invasion (Atayde et al 2004). Quantitative RT-PCR and dot-blot hybridisation experiments have shown that metacyclic trypomastigotes upregulate gp82 mRNA (Songthamwat et al 2007, Gentil et al 2009). In contrast to nuclear run-on assays, which showed that the transcriptional rates of gp82 genes were similar between epimastigotes and metacyclic trypomastigotes, northern blot assays showed that the half-life of gp82 mRNA is dramatically reduced in epimastigotes.…”

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confidence: 83%

Regulatory elements involved in the post-transcriptional control of stage-specific gene expression in Trypanosoma cruzi: a review

Araújo

Teixeira

2011

Mem. Inst. Oswaldo Cruz

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Trypanosoma cruzi is an intracellular parasite that is transmitted by at least 40 different blood-sucking triatomine species to over 1,000 mammalian species (Brener et al. 2000). This parasite must adapt to enormous changes in its extracellular milieu, such as changes in environmental temperature as well as in the available nutrients inside each host. The parasite has a complex life cycle that is characterised by four stages; epimastigotes and metacyclic trypomastigotes are present in the insect vector, whereas intracellular amastigotes and bloodstream trypomastigotes are present in the mammalian host. Thus, this species must develop a broad set of molecular tools that allow it to multiply in the insect gut, to invade and multiply inside a large number of distinct mammalian cell types and to circumvent host immune defence systems. To meet such phenotypic plasticity, T. cruzi relies on unique mechanisms that can control the expression of its repertoire of about 12,000 genes. Because its genome is constitutively transcribed into long polycistronic primary transcripts, mRNAs for proteincoding genes must be processed through trans-splicing and polyadenylation reactions. The mRNAs must also interact with different protein factors in a complex posttranscriptional regulatory machinery that determines the levels of their protein product according to the cellular demands of the parasite in each stage of its life cycle.In the same issue that the T. cruzi genome was published (El-Sayed et al. 2005a), a study describing its proteome was also reported (Atwood et al. 2005). Proteins extracted from whole-cell and subcellular lysates of the four stages of T. cruzi were analysed by mass spectrometry (epimastigotes, metacyclic trypomastigotes, amastigotes and trypomastigotes), which identified 2,784 proteins belonging to the 1,168 protein groups in the annotated T. cruzi genome. Although about 30% of the identified proteins were found at all life-cycle stages, at least 248 proteins were only expressed at one stage, thereby demonstrating significant changes in the relative abundance of T. cruzi proteins throughout its life cycle. One of the main findings in these proteomic analyses was that the four parasite stages use distinct energy sources (Atwood et al. 2005); intracellular amastigotes upregulated proteins involved in lipid-dependent energy metabolism, such as enzymes of the citric acid cycle, whereas enzymes capable of catalysing the conversion of histidine to glutamate were more abundant in the insect epimastigote stage. Heat-shock proteins (HSP) and proteins involved in vesicular trafficking were also preferentially detected in amastigotes. Furthermore, enzymes involved in antioxidant defence were upregulated during the transformation of epimastigotes into invasive metacyclic trypomastigotes, whereas bloodstream trypomastigotes upregulated the surface expression of several large gene families that are known to be involved in interacting with the mammalian host (Atwood et al. 2005).In agreement with this proteomics data, glo...

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“…Steady-state levels of GP82 transcripts from T. cruzi G strain were determined by northern blot, dot-blot hybridization, and quantitative real-time PCR, demonstrating that there is a significant increase in GP82 mRNA levels in metacyclic forms when compared with the other three stages [9, 11, 60]. Northern blot analysis revealed a single band of 2.2 kb mRNA only in metacyclic forms [9].…”

Section: Post-transcriptional Control Mechanisms Of Gp82 Expressionmentioning

confidence: 99%

“…Similar results were obtained using quantitative real-time PCR (unpublished data). Moreover, expression analysis of other three GP82 gene subfamilies from Peru-2 strain, called groups A, B, and C, showed an increase in mRNA accumulation (4.7 to 9.3-fold) in metacyclic forms when compared to epimastigotes [11]. Additionally, GP82 protein stage-specific expression was also showed by western blot using the Mab3F6 [3].…”

Section: Post-transcriptional Control Mechanisms Of Gp82 Expressionmentioning

confidence: 99%

Genetic Structure and Expression of the Surface Glycoprotein GP82, the Main Adhesin ofTrypanosoma cruziMetacyclic Trypomastigotes

Corrêa

Cordero

Gentil

et al. 2013

The Scientific World Journal

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T. cruzi improves the likelihood of invading or adapting to the host through its capacity to present a large repertoire of surface molecules. The metacyclic stage-specific surface glycoprotein GP82 has been implicated in host cell invasion. GP82 is encoded by multiple genes from the trans-sialidase superfamily. GP82 shows a modular organization, with some variation of N-terminal region flanking a conserved central core where the binding sites to the mammalian cell and gastric mucin are located. The function of GP82 as adhesin in host cell invasion process could expose the protein to an intense conservative and selective pressure. GP82 is a GPI-anchored surface protein, synthesized as a 70 kDa precursor devoid of N-linked sugars. GPI-minus variants accumulate in the ER indicating that GPI anchor acts as a forward transport signal for progressing along the secretory pathway as suggested for T. cruzi mucins. It has been demonstrated that the expression of GP82 is constitutive and may be regulated at post-transcriptional level, for instance, at translational level and/or mRNA stabilization. GP82 mRNAs are mobilized to polysomes and consequently translated, but only in metacyclic trypomastigotes. Analysis of transgenic parasites indicates that the mechanism regulating GP82 expression involves multiple elements in the 3′UTR.

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Structure and expression of three gp82 gene subfamilies of Trypanosoma cruzi

Cited by 6 publications

References 38 publications

Lineage Analysis of Circulating Trypanosoma cruzi Parasites and Their Association with Clinical Forms of Chagas Disease in Bolivia

Lineage Analysis of Circulating Trypanosoma cruzi Parasites and Their Association with Clinical Forms of Chagas Disease in Bolivia

Regulatory elements involved in the post-transcriptional control of stage-specific gene expression in Trypanosoma cruzi: a review

Genetic Structure and Expression of the Surface Glycoprotein GP82, the Main Adhesin ofTrypanosoma cruziMetacyclic Trypomastigotes

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