Downregulation of gamma subunit of TCP1 chaperonin of Leishmania donovani modulates extracellular vesicles-mediated macrophage microbicidal function.

Shailendra Yadav, Apeksha Anand, Deep Chandra Balodi, Karthik Ramalingam, Kalyan Mitra, Jaswinder Singh Maras, Neena Goyal

Microbial pathogenesis 2022 Aug

filter terms:

T-complex protein-1 (TCP1) is a group II chaperonin, known to fold various proteins like actin and tubulin. In Leishmania donovani only one subunit that is gamma subunit (LdTCP1γ) has been functionally characterized as a homo-oligomeric complex that exhibits ATP-dependent protein folding. The gene is essential for the survival and infectivity of the parasite. Leishmania parasite releases extracellular vesicles (EVs) containing numerous virulence factors, which play an essential role in parasite pathogenesis and modulate host immune cell signaling. The present study demonstrates that LdTCP1γ is secreted in the EVs and modulates host macrophage functions. EVs isolated from LdTCP1γ single-allele-replacement mutants significantly upregulate the microbicidal function of LPS-induced macrophage as evident by increased levels of proinflammatory cytokines (TNF-α, IL-6), iNOS and NO production. Further, the comparative proteomics of wild-type and single-allele-replacement mutant EVs showed that out of 876 identified proteins, 207 were significantly modulated. Among them, the top 50 modulated and abundantly secreted proteins constitute ∼40% of the total identified protein intensity and include virulence factors such as GP63, peroxiredoxin, enolase, HSP70, elongation factor 2, amastin, eukaryotic translation initiation factor and α-tubulin. The comparative proteomic analysis revealed that the proteome enrichment of the EVs from LdTCP1γ single-allele replacement mutants significantly differs from wild-type EVs, which may be responsible for the altered host microbicidal responses. Thus, our data provide new insight into the role of LdTCP1γ in EVs-mediated host-parasite interactions. Copyright © 2022 Elsevier Ltd. All rights reserved.

Citation

Shailendra Yadav, Apeksha Anand, Deep Chandra Balodi, Karthik Ramalingam, Kalyan Mitra, Jaswinder Singh Maras, Neena Goyal. Downregulation of gamma subunit of TCP1 chaperonin of Leishmania donovani modulates extracellular vesicles-mediated macrophage microbicidal function. Microbial pathogenesis. 2022 Aug;169:105616