TransLeish: Identification of membrane transporters essential for survival of intracellular Leishmania parasites in a systematic gene deletion screen
Abstract 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 transport...
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Nature Portfolio
2025-01-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-024-55538-7 |
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| author | Andreia Albuquerque-Wendt Ciaran McCoy Rachel Neish Ulrich Dobramysl Çağla Alagöz Tom Beneke Sally A. Cowley Kathryn Crouch Richard J. Wheeler Jeremy C. Mottram Eva Gluenz |
| author_facet | Andreia Albuquerque-Wendt Ciaran McCoy Rachel Neish Ulrich Dobramysl Çağla Alagöz Tom Beneke Sally A. Cowley Kathryn Crouch Richard J. Wheeler Jeremy C. Mottram Eva Gluenz |
| author_sort | Andreia Albuquerque-Wendt |
| collection | DOAJ |
| description | Abstract 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. |
| format | Article |
| id | doaj-art-c839ea437a5a40608fec2ad500cb81e6 |
| institution | DOAJ |
| issn | 2041-1723 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj-art-c839ea437a5a40608fec2ad500cb81e62025-08-20T02:53:46ZengNature PortfolioNature Communications2041-17232025-01-0116111710.1038/s41467-024-55538-7TransLeish: Identification of membrane transporters essential for survival of intracellular Leishmania parasites in a systematic gene deletion screenAndreia Albuquerque-Wendt0Ciaran McCoy1Rachel Neish2Ulrich Dobramysl3Çağla Alagöz4Tom Beneke5Sally A. Cowley6Kathryn Crouch7Richard J. Wheeler8Jeremy C. Mottram9Eva Gluenz10School of Infection and Immunity, University of Glasgow, Sir Graeme Davies Building, 120 University PlaceUniversity of Oxford, Sir William Dunn School of Pathology, South Parks RoadYork Biomedical Research Institute, Department of Biology, University of YorkMedawar Building for Pathogen Research, Nuffield Department of Medicine, University of OxfordInstitute of Cell Biology, University of Bern, Baltzerstrasse 4University of Oxford, Sir William Dunn School of Pathology, South Parks RoadJames and Lillian Martin Centre for Stem Cell Research, Sir William Dunn School of Pathology, University of OxfordSchool of Infection and Immunity, University of Glasgow, Sir Graeme Davies Building, 120 University PlaceMedawar Building for Pathogen Research, Nuffield Department of Medicine, University of OxfordYork Biomedical Research Institute, Department of Biology, University of YorkSchool of Infection and Immunity, University of Glasgow, Sir Graeme Davies Building, 120 University PlaceAbstract 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.https://doi.org/10.1038/s41467-024-55538-7 |
| spellingShingle | Andreia Albuquerque-Wendt Ciaran McCoy Rachel Neish Ulrich Dobramysl Çağla Alagöz Tom Beneke Sally A. Cowley Kathryn Crouch Richard J. Wheeler Jeremy C. Mottram Eva Gluenz TransLeish: Identification of membrane transporters essential for survival of intracellular Leishmania parasites in a systematic gene deletion screen Nature Communications |
| title | TransLeish: Identification of membrane transporters essential for survival of intracellular Leishmania parasites in a systematic gene deletion screen |
| title_full | TransLeish: Identification of membrane transporters essential for survival of intracellular Leishmania parasites in a systematic gene deletion screen |
| title_fullStr | TransLeish: Identification of membrane transporters essential for survival of intracellular Leishmania parasites in a systematic gene deletion screen |
| title_full_unstemmed | TransLeish: Identification of membrane transporters essential for survival of intracellular Leishmania parasites in a systematic gene deletion screen |
| title_short | TransLeish: Identification of membrane transporters essential for survival of intracellular Leishmania parasites in a systematic gene deletion screen |
| title_sort | transleish identification of membrane transporters essential for survival of intracellular leishmania parasites in a systematic gene deletion screen |
| url | https://doi.org/10.1038/s41467-024-55538-7 |
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