Deletion of a Golgi protein in Trypanosoma cruzi reveals a critical role for Mn2+ in protein glycosylation needed for host cell invasion and intracellular replication

Abstract

Trypanosoma cruzi is a protist parasite and the causative agent of American trypanosomiasis or Chagas disease. The parasite life cycle in its mammalian host includes an intracellular stage, and glycosylated proteins play a key role in host-parasite interaction facilitating adhesion, invasion and immune evasion. Here, we report that a Golgi-localized Mn2+-Ca2+/H+ exchanger of T. cruzi (TcGDT1) is required for efficient protein glycosylation, host cell invasion, and intracellular replication. The Golgi localization was determined by immunofluorescence and electron microscopy assays. TcGDT1 was able to complement the growth defect of Saccharomyces cerevisiae null mutants of its ortholog ScGDT1 but ablation of TcGDT1 by CRISPR/Cas9 did not affect the growth of the insect stage of the parasite. The defect in protein glycosylation was rescued by Mn2+ supplementation to the growth medium, underscoring the importance of this transition metal for Golgi glycosylation of proteins.

The Development of Aspergillus flavus and Biosynthesis of Aflatoxin B1 are Regulated by the Golgi-Localized Mn2+ Transporter Pmr1

“…cerevisiae, Trypanosoma cruzi, and A. thaliana. ,, Accordingly, Gdt1 may function as a complementary factor in A. flavus, mitigating growth deficiencies caused by Pmr1 deficits.…”

“…Moreover, our investigation unveils that Pmr1 is a Ca 2+ and Mn 2+ transporter localized to the Golgi apparatus, where proteins are processed, sorted, and transported. Previous research has indicated the participation of both Pmr1 and Gdt1 in diverse organisms, including yeast, parasites, and plants regarding Ca 2+ - and Mn 2+ -dependent protein glycosylation processes. ,, The process of glycosylation is widely recognized as one of the most prevalent peptide modifications in nature, with over two-thirds of eukaryotic proteins undergoing this modification . Consequently, we attempt to investigate whether the addition of exogenous Mn 2+ could potentially restore the homeostasis of protein glycosylation in the Δ pmr1 mutant.…”

“…Gdt1, a member of the UPF0016 protein family, has been experimentally confirmed as a Golgi-localized protein responsible for maintaining Mn 2+ homeostasis across S. cerevisiae, Trypanosoma cruzi, and A. thaliana. 14,15,51 Accordingly, Gdt1 may function as a complementary factor in A. flavus, mitigating growth deficiencies caused by Pmr1 deficits. Surprisingly, qPCR analysis has revealed a significant decrease in gdt1 mRNA expression within the Δpmr1 mutant strain.…”

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.…”