Characterization of a Thioredoxin-1 Gene from<i>Taenia solium</i>and Its Encoding Product

Jiménez, Lucı́a; Rodríguez-Lima, Oscar; Ochoa‐Sánchez, Alicia; Landa, Abraham

doi:10.1155/2015/453469

Cited by 7 publications

(3 citation statements)

References 35 publications

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“…In the proximal promoter gene, we found putative binding sites for several general transcription factors related to the formation of transcription preinitiation complex, which were all localized at appropriate distances to be functional [ 46 , 47 ]. These putative sites have been reported previously in T. solium genes [ 48 , 49 , 50 , 51 ]. In addition, the presence of regulator elements such as XRE, AhR, ARNT, and AP‐1 were identified in the distal promoter of Ts24gst , which were are also found in promoters of GST genes in E. granulosus , as well as other GST and detoxifying enzyme genes [ 26 , 52 , 53 ].…”

Section: Discussionsupporting

confidence: 81%

Biochemical characterization and gene structure analysis of the 24‐kDa glutathione transferase sigma from Taenia solium

Miranda‐Blancas,

Rodríguez‐Lima,

García‐Gutiérrez

et al. 2024

FEBS Open Bio

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Taenia solium can cause human taeniasis and/or cysticercosis. The latter can in some instances cause human neurocysticercosis which is considered a priority in disease‐control strategies and the prevention of mental health problems. Glutathione transferases are crucial for the establishment and long‐term survival of T. solium; therefore, we structurally analyzed the 24‐kDa glutathione transferase gene (Ts24gst) of T. solium and biochemically characterized its product. The gene promoter showed potential binding sites for transcription factors and xenobiotic regulatory elements. The gene consists of a transcription start site, four exons split by three introns, and a polyadenylation site. The gene architecture is conserved in cestodes. Recombinant Ts24GST (rTs24GST) was active and dimeric. Anti‐rTs24GST serum showed slight cross‐reactivity with human sigma‐class GST. A 3D model of Ts24GST enabled identification of putative residues involved in interactions of the G‐site with GSH and of the H‐site with CDNB and prostaglandin D2. Furthermore, rTs24GST showed optimal activity at 45 °C and pH 9, as well as high structural stability in a wide range of temperatures and pHs. These results contribute to the better understanding of this parasite and the efforts directed to fight taeniasis/cysticercosis.

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Section: Discussionsupporting

confidence: 81%

Biochemical characterization and gene structure analysis of the 24‐kDa glutathione transferase sigma from Taenia solium

Miranda‐Blancas,

Rodríguez‐Lima,

García‐Gutiérrez

et al. 2024

FEBS Open Bio

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“…Analysis of regulatory elements presents in core promoters of Taeniids genes. A) Alignment of core promoter sequences from genes of: Actin 5 (pAT5, [35]), Actin 6 (pAT6, [35]), 2-Cys Peroxiredoxin (TsPrx, [35]), Thioredoxin-1 (TsTrx1, [36]), Cu,Zn Superoxide dismutase (TsCu,ZnSOD, [37]) and TBP1 of T. solium (TsTBP1); 2-Cys Peroxiredoxin of Taenia crassiceps (TcPrx, [35]); and Actin I (EgactI, [38]), Actin II (EgactII, [38]), gen Homebox-containig 1 (EgHbx1, [39]) of Echinococcus granulosus. -TAF6 antibody ----+ ----Anti-TAF9 antibody -----+ ---Consensus DPE ------+ --Consensus DPE mut -------+ -TsTBP1-DPEmut probe --------+ 1 2 3 4 5 6 A B C D Figure 10…”

Section: Relatives To Tss Showing a Consensus Sequence T-a-t-a-t/a-tmentioning

confidence: 99%

Taenia soliumTAF6 and TAF9 binds to a putative Downstream Promoter Element present inTsTBP1gene core promoter

Rodríguez-Lima

García-Gutiérrez

Jiménez

et al. 2017

Preprint

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Edificio A 2do piso. Ciudad Universitaria. CDMX 04510, México Phone: (52 55) 56-23-23-57, Fax: (52 55) 56-23-23-82, Email: landap@unam.mx 2 Abstract.We have cloned and characterized the gene encoding to Taenia solium TATA binding protein 1 (TsTBP1). It spans 1481 bp and its coding region is interrupted by four introns that possess the consensus donor/acceptor sequences. It produces a protein of 238 amino acids residues, which presented all the classical motives of the TBP1. On the core promoter region we identified putative binding sites for NF1, AP-1, YY1, TAF1/TAF2 and TAF6/TAF9, and the TSS that corresponds to an A +1 . Southern and Northern blot analysis showed that TsTBP1 is encoded by a single gene, which produce a messenger of about 1.1 kbp with a higher differential expression in adult than the larval stage. Moreover, two putative TATA boxes were identified at -97 and -69 bp (relative to the TSS), likewise a Downstream Promoter Element (DPE) was located at +27 to +31 bp. EMSA experiments did not show any component of cysticerci nuclear extracts bound to the putative TATA-box elements; in contrast TAF6 and TAF9 bind to the DPE, showing that TsTBP1 is a TATAless gen. On the other hand, TAF6 and TAF9 were localized on the nucleus from cells of T. crassiceps cysticerci bladder walls. By using the amino acid sequences of T. solium TAF6 (TsTAF6) and TAF9 (TsTAF9), we constructed a molecular model showing interaction between DPE-TsTAF6 and TsTAF6-TsTAF9. Finally, an in silico analysis of the core promoters of Taeniidae family genes showed that TATA-box and DPE are homologous to the mammalian elements, but not so the Inr. The low identity between TAF9 from cestodes and mammals open the possibility to use it as target to interrupt or modulate the transcription of TATA-box less genes in cestodes as a novel therapeutic strategy. 3 Author summary. Neurocysticercosis still being a health problem in developing countries. Mexico reports a prevalence of 2.5% in the total patients from National Institute of Neurology and Neurosurgery. Several efforts have been made in different fields to study Taenia solium, and although the Genome Project has been published and released the genomic sequence of this parasite; the transcriptional mechanisms this organism remains unexplored. We isolated and characterized Taenia solium TATA-Binding Protein 1 (TsTBP1) gene and TBP-associated factor 6 and 9 cDNAs (TsTAF6 and TsTAF9, respectively). Our principal findings are: 1.-Identification of cis and trans elements in the core promoter of TsTBP1 gene. 2.-The TsTBP1 gene is a TATA-less promoter. 3.-Cloning of cDNAs to TsTAF6 and TsTAF9, 4.-Identification of a DPE in the core promoter of TsTBP1 gene, which interacts with TAF6 and TAF9, 5.-Construction of a molecular model that shows the interaction between DPE, TAF6, and TAF9; and 6.-A proposal of consensus sequences for TATAbox, Inr and DPE presented on Taeniidae family core promoters.4

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“…Although few analyses for expression stability of RG have been done in other cestodes 24,25 , there is no report of a similar validation for the pre-adult stages of T. solium . Data normalization for RT-qPCR comparisons were performed previously, however, using canonical housekeeping gene targets such as glyceraldehyde-3-phosphate dehydrogenase (GAPDH) 26 or Cu/Zn superoxide dismutase (TsCu/ZnSOD) 27 .…”

Section: Introductionmentioning

confidence: 99%

Validation of Reference Genes for RT-qPCR Relative Expression Analysis in Pre-Adult Stages ofTaenia solium

Gutiérrez-Loli

Torre

Aguila

et al. 2022

Preprint

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SUMMARYThe larvae-to-adult development on the life cycle of zoonotic parasitic tapewormTaenia soliuminvolves striking -but clinically unappreciated-events with pivotal importance in cestode biology. Unlike the ones related to the intermediate host, the early-adult stages can be addressedin vitrooffering a useful model to study evagination, strobilation and worm development. In the absence of a stage-specific transcriptome, postgenomic data exploration followed by single-gene relative expression analysis by RT-qPCR (reverse transcription-quantitative PCR) are useful strategies to gather information on the regulation of genes of interest during parasite development. However, this approach requires the validation of an endogenous reference gene (RG) to achieve accurate comparisons.Therefore, we analyzed the expression stability of 17 candidate RGs on the context of the early-adult stages ofT. soliumclassified as non-evaginated and evaginated larvae (cysts). The comprehensive tool RefFinder ranked malate dehydrogenase as the most stable gene within these conditions, and its suitability for relative quantification was validated by normalizing the expression of the transporter TGTP1 gene, known for being upregulated upon evagination. This study is the first attempt in finding reliable normalization standards for transcript exploration in genus Taenia.

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Characterization of a Thioredoxin-1 Gene fromTaenia soliumand Its Encoding Product

Cited by 7 publications

References 35 publications

Biochemical characterization and gene structure analysis of the 24‐kDa glutathione transferase sigma from Taenia solium

Biochemical characterization and gene structure analysis of the 24‐kDa glutathione transferase sigma from Taenia solium

Taenia soliumTAF6 and TAF9 binds to a putative Downstream Promoter Element present inTsTBP1gene core promoter

Validation of Reference Genes for RT-qPCR Relative Expression Analysis in Pre-Adult Stages ofTaenia solium

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