TransLeish: Identification of membrane transporters essential for survival of intracellular Leishmania parasites in a systematic gene deletion screen

TY - JOUR

T1 - TransLeish

T2 - Identification of membrane transporters essential for survival of intracellular Leishmania parasites in a systematic gene deletion screen

AU - Albuquerque-Wendt, Andreia

AU - McCoy, Ciaran

AU - Neish, Rachel

AU - Dobramysl, Ulrich

AU - Alagöz, Çağla

AU - Beneke, Tom

AU - Cowley, Sally A

AU - Crouch, Kathryn

AU - Wheeler, Richard J

AU - Mottram, Jeremy C

AU - Gluenz, Eva

N1 - © 2024. The Author(s).

PY - 2025/1/2

Y1 - 2025/1/2

N2 - For the protozoan parasite Leishmania, completion of its life cycle requires sequential adaptation of cellular physiology and nutrient scavenging mechanisms to the different environments of a sand fly alimentary tract and the acidic mammalian host cell phagolysosome. Transmembrane transporters are the gatekeepers of intracellular environments, controlling the flux of solutes and ions across membranes. To discover which transporters are vital for survival as intracellular amastigote forms, we carried out a systematic loss-of-function screen of the L. mexicana transportome. A total of 312 protein components of small molecule carriers, ion channels and pumps were identified and targeted in a CRISPR-Cas9 gene deletion screen in the promastigote form, yielding 188 viable null mutants. Forty transporter deletions caused significant loss of fitness in macrophage and mouse infections. A striking example is the Vacuolar H+ ATPase (V-ATPase), which, unexpectedly, was dispensable for promastigote growth in vitro but essential for survival of the disease-causing amastigotes.

AB - For the protozoan parasite Leishmania, completion of its life cycle requires sequential adaptation of cellular physiology and nutrient scavenging mechanisms to the different environments of a sand fly alimentary tract and the acidic mammalian host cell phagolysosome. Transmembrane transporters are the gatekeepers of intracellular environments, controlling the flux of solutes and ions across membranes. To discover which transporters are vital for survival as intracellular amastigote forms, we carried out a systematic loss-of-function screen of the L. mexicana transportome. A total of 312 protein components of small molecule carriers, ion channels and pumps were identified and targeted in a CRISPR-Cas9 gene deletion screen in the promastigote form, yielding 188 viable null mutants. Forty transporter deletions caused significant loss of fitness in macrophage and mouse infections. A striking example is the Vacuolar H+ ATPase (V-ATPase), which, unexpectedly, was dispensable for promastigote growth in vitro but essential for survival of the disease-causing amastigotes.

KW - Animals

KW - Mice

KW - Gene Deletion

KW - Macrophages/parasitology

KW - CRISPR-Cas Systems

KW - Protozoan Proteins/genetics

KW - Membrane Transport Proteins/genetics

KW - Leishmania mexicana/genetics

KW - Vacuolar Proton-Translocating ATPases/genetics

KW - Female

KW - Mice, Inbred BALB C

U2 - 10.1038/s41467-024-55538-7

DO - 10.1038/s41467-024-55538-7

M3 - Article

C2 - 39747086

SN - 2041-1723

VL - 16

JO - Nature Communications

JF - Nature Communications

M1 - 299

ER -