Trypanosoma cruzi has Two Peptidyl-tRNA Hydrolases Showing Different Localization and Function

IF 1.2 3区 农林科学 Q4 PARASITOLOGY
María Elizabeth Reséndiz-Juárez, Ana Laura Rosas-Soto, Armando Pérez-Rangel, José Tapia-Ramírez, Emmanuel Ríos-Castro, Fanny Rodríguez-Cruz, Ricardo Alejandre-Aguilar, Rebeca Manning-Cela, Gloria León-Avila, José Manuel Hernández-Hernández
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Abstract

Purpose

Peptidyl-tRNA hydrolase (Pth), first described in Escherichia coli, is responsible for rescuing stalled ribosomes during peptidyl-tRNA “drop off”. Bacterial Pth has been widely studied, but the characterization of eukaryotic Pth remains a poorly researched field, especially in protozoan parasites. This work aimed to characterize Trypanosoma cruzi Pths and determine their localization.

Methods

Two open reading frames (ORFs) that may encode Pths were identified in the T. cruzi genome. Bioinformatics analysis was performed for each protein using conserved domain analysis and multiple alignment. ORFs were cloned into an expression vector, E. coli pth(Ts) competent cells were transformed, and thermosensitivity tests were performed. Recombinant proteins were expressed and purified to immunize rats and obtain polyclonal antibodies. Pull down and immunoprecipitation followed by mass spectrometry to verify the interactions.

Results

TcPth and TcPth2 have a conserved domain corresponding to the Pth2 superfamily. Multiple alignments with previously characterized amino acid sequences of Pths showed that they are unrelated to T. cruzi proteins, considering that conserved residues of catalytic importance are absent. TcPth was able to rescue the E. coli thermosensitive pth(Ts) mutation, but TcPth2 was not. TcPth2 interacts with reservosome proteins such as cysteine peptidase and endocytic pathway proteins.

Conclusion

The results suggest that TcPth and TcPth2 has a different function. This work represents the first in its area since the Pths of the T. cruzi were characterized and breaks ground for the characterization of Pths from other protozoan parasites.

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来源期刊
Acta Parasitologica
Acta Parasitologica 医学-寄生虫学
CiteScore
3.10
自引率
6.70%
发文量
149
审稿时长
6-12 weeks
期刊介绍: Acta Parasitologica is an international journal covering the latest advances in the subject. Acta Parasitologica publishes original papers on all aspects of parasitology and host-parasite relationships, including the latest discoveries in biochemical and molecular biology of parasites, their physiology, morphology, taxonomy and ecology, as well as original research papers on immunology, pathology, and epidemiology of parasitic diseases in the context of medical, veterinary and biological sciences. The journal also publishes short research notes, invited review articles, book reviews. The journal was founded in 1953 as "Acta Parasitologica Polonica" by the Polish Parasitological Society and since 1954 has been published by W. Stefanski Institute of Parasitology of the Polish Academy of Sciences in Warsaw. Since 1992 in has appeared as Acta Parasitologica in four issues per year.
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