Characterization and functional analysis of the proteins Prohibitin 1 and 2 in Trypanosoma cruzi

PLoS Negl Trop Dis. 2021 Apr 8;15(4):e0009322. doi: 10.1371/journal.pntd.0009322. eCollection 2021 Apr.

Abstract

Background: Chagas disease is the third most important neglected tropical disease. There is no vaccine available, and only two drugs are generally prescribed for the treatment, both of which with a wide range of side effects. Our study of T. cruzi PHBs revealed a pleiotropic function in different stages of the parasite, participating actively in the transformation of the non-infective replicative epimastigote form into metacyclic trypomastigotes and also in the multiplication of intracellular amastigotes.

Methodology/principal findings: To obtain and confirm our results, we applied several tools and techniques such as electron microscopy, immuno-electron microscopy, bioinformatics analysis and molecular biology. We transfected T. cruzi clones with the PHB genes, in order to overexpress the proteins and performed a CRISPR/Cas9 disruption to obtain partially silenced PHB1 parasites or completely silenced PHB2 parasites. The function of these proteins was also studied in the biology of the parasite, specifically in the transformation rate from non-infective forms to the metacyclic infective forms, and in their capacity of intracellular multiplication.

Conclusion/significance: This research expands our understanding of the functions of PHBs in the life cycle of the parasite. It also highlights the protective role of prohibitins against ROS and reveals that the absence of PHB2 has a lethal effect on the parasite, a fact that could support the consideration of this protein as a possible target for therapeutic action.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Chagas Disease / parasitology*
  • Clustered Regularly Interspaced Short Palindromic Repeats
  • Computer Simulation
  • Life Cycle Stages*
  • Male
  • Mice
  • Mice, Inbred BALB C
  • Microscopy, Electron, Scanning
  • Prohibitins
  • Protozoan Proteins / analysis
  • Rats
  • Rats, Wistar
  • Repressor Proteins / genetics
  • Repressor Proteins / metabolism*
  • Trypanosoma cruzi / enzymology*
  • Trypanosoma cruzi / genetics

Substances

  • Phb protein, rat
  • Phb2 protein, mouse
  • Prohibitins
  • Protozoan Proteins
  • Repressor Proteins

Grants and funding

This work was funded through grants awarded to AO from Instituto Carlos III, Ministerio de Sanidad of the Government of Spain by the UE ERANet program: “Research in prevention of Congenital Chagas Disease Parasitological, placental and immunological markers (Cochaco)” (ERANet17/HLH-0142); from Fundación Ramón Areces: “Interactoma de las Exovesículas de T. cruzi y de los inmunocomplejos que forman con las células del hospedador; Implicaciones en la patología de la Enfermedad de Chagas”; from Ministerio de Ciencia y Tecnología of the Government of Spain: “Exovesicles of Trypanosoma cruzi and the immunocomplexes that they form. Implications in the pathology of Chagas disease (ExCha)” (PGC2018-099424-B-I00). In addition, AGS is financially supported by the UE ERANet program: “Research in prevention of congenital Chagas disease: Parasitological, placental and immunological markers” (ELAC2014/HID-¬‐0328. ERANET UE ERANE-LAC HD 328/2014). AKI-V was supported by ITAIPU Binational and BECAL of the Government of Paraguay, and she was a doctoral student in the Biomedicine Program at University of Granada (Spain). LR-M was supported by the Oficina de Asuntos Internacionales y Cooperación Externa (OAICE) of the Universidad de Costa Rica and the Ministerio de Ciencia, Tecnología y Telecomunicaciones (MICITT) of the Government of Costa Rica. And AC-G was suported by the “Beca de Iniciación a la Investigación para estudiantes de Máster”, Universidad de Granada (Spain). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.