Genetic Validation of Leishmania Genes Essential for Amastigote Survival in Vivo Using N-myristoyltransferase As a Model

Overview

Journal Parasit Vectors

Publisher Biomed Central

Specialties Parasitology
Tropical Medicine

Date 2020 Mar 16

PMID 32171322

Citations 6

Authors

Daniel Paape

Catriona T Prendergast

Helen P Price

Johannes S P Doehl

Deborah F Smith

Affiliations

Soon will be listed here.

Abstract

Background: Proving that specific genes are essential for the intracellular viability of Leishmania parasites within macrophages remains a challenge for the identification of suitable targets for drug development. This is especially evident in the absence of a robust inducible expression system or functioning RNAi machinery that works in all Leishmania species. Currently, if a target gene of interest in extracellular parasites can only be deleted from its genomic locus in the presence of ectopic expression from a wild type copy, it is assumed that this gene will also be essential for viability in disease-promoting intracellular parasites. However, functional essentiality must be proven independently in both life-cycle stages for robust validation of the gene of interest as a putative target for chemical intervention.

Methods: Here, we have used plasmid shuffle methods in vivo to provide supportive genetic evidence that N-myristoyltransferase (NMT) is essential for Leishmania viability throughout the parasite life-cycle. Following confirmation of NMT essentiality in vector-transmitted promastigotes, a range of mutant parasites were used to infect mice prior to negative selection pressure to test the hypothesis that NMT is also essential for parasite viability in an established infection.

Results: Ectopically-expressed NMT was only dispensable under negative selection in the presence of another copy. Total parasite burdens in animals subjected to negative selection were comparable to control groups only if an additional NMT copy, not affected by the negative selection, was expressed.

Conclusions: NMT is an essential gene in all parasite life-cycle stages, confirming its role as a genetically-validated target for drug development.

Citing Articles

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Comparative Proteomics and Genome-Wide Druggability Analyses Prioritized Promising Therapeutic Targets against Drug-Resistant .

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Expanded Proteomic Survey of the Human Parasite Leishmania major Focusing on Changes in Null Mutants of the Golgi GDP-Mannose/Fucose/Arabinopyranose Transporter and of the Mitochondrial Fucosyltransferase .

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The Golgi-localized sphingosine-1-phosphate phosphatase is indispensable for Leishmania major.

Okundaye B, Biyani N, Moitra S, Zhang K Sci Rep. 2022; 12(1):16064.

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Brannigan J, Wilkinson A Biophys Rev. 2022; 13(6):1139-1146.

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