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

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.

Deep Learning-based structural and functional annotation of Pandoravirus hypothetical proteins

“…FoldSeek recovered the same similarities to stomatins and flotillins in the search against the PDB but gave stronger hits to AFDB entries for prohibitins. Prohibitins, stomatins and flotillins all share a conserved domain, namely the Band 7/SPFH/PH b -domain containing proteins (Morrow and Parton, 2005; Ibarrola-Vannucci et al ., 2021). Intriguingly, prohibitins localise to the mitochondrial membrane, matching the Deeploc predicted localization for cluster 8.…”

The Elusive Trypanosoma cruzi Disperse Gene Protein Family (DGF-1)

“…In most assays focused on the detection of RNAs by qPCR and proteins using specific antibodies, there is better agreement [ 13 , 15 , 45 , 46 , 52 ] than in whole proteome and transcriptome techniques. Most proteomic techniques will underestimate or miss proteins with modifications like lipidation, glycosylation, etc., and in silico prediction reveals that DGF-1 has many of these modification sites.…”

Ablation of the P21 Gene of Trypanosoma cruzi Provides Evidence of P21 as a Mediator in the Control of Epimastigote and Intracellular Amastigote Replication

“…For protozoan parasites, the CRISPR/Cas9 system provides the opportunity to gain new insights into parasite biology and allows the development of new approaches to study pathogenesis and parasite-host interactions at the cellular and molecular levels (Zhang et al, 2014;Sollelis et al, 2015;Lander et al, 2016;Sidik et al, 2016;Young et al, 2019). Several studies have used these tools to deepen the functional understanding of T. cruzi and its interaction with host cells (Peng et al, 2014;Lander et al, 2015;Chiurillo et al, 2017;Lander et al, 2017;Costa et al, 2018;Akutsu et al, 2019;Chiurillo et al, 2019;Tavernelli et al, 2019;Chiurillo et al, 2020;Lander et al, 2020;Costa-Silva et al, 2021;Ibarrola-Vannucci et al, 2021, Ramakrishnan et al, 2021.…”