The <italic toggle="yes">Plasmodium falciparum</italic> Artemisinin Susceptibility-Associated AP-2 Adaptin μ Subunit is Clathrin Independent and Essential for Schizont Maturation
ABSTRACT The efficacy of current antimalarial drugs is threatened by reduced susceptibility of Plasmodium falciparum to artemisinin, associated with mutations in pfkelch13. Another gene with variants known to modulate the response to artemisinin encodes the μ subunit of the AP-2 adaptin trafficking...
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| Format: | Article |
| Language: | English |
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American Society for Microbiology
2020-02-01
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| Series: | mBio |
| Subjects: | |
| Online Access: | https://journals.asm.org/doi/10.1128/mBio.02918-19 |
| _version_ | 1818740173373964288 |
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| author | Ryan C. Henrici Rachel L. Edwards Martin Zoltner Donelly A. van Schalkwyk Melissa N. Hart Franziska Mohring Robert W. Moon Stephanie D. Nofal Avnish Patel Christian Flueck David A. Baker Audrey R. Odom John Mark C. Field Colin J. Sutherland |
| author_facet | Ryan C. Henrici Rachel L. Edwards Martin Zoltner Donelly A. van Schalkwyk Melissa N. Hart Franziska Mohring Robert W. Moon Stephanie D. Nofal Avnish Patel Christian Flueck David A. Baker Audrey R. Odom John Mark C. Field Colin J. Sutherland |
| author_sort | Ryan C. Henrici |
| collection | DOAJ |
| description | ABSTRACT The efficacy of current antimalarial drugs is threatened by reduced susceptibility of Plasmodium falciparum to artemisinin, associated with mutations in pfkelch13. Another gene with variants known to modulate the response to artemisinin encodes the μ subunit of the AP-2 adaptin trafficking complex. To elucidate the cellular role of AP-2μ in P. falciparum, we performed a conditional gene knockout, which severely disrupted schizont organization and maturation, leading to mislocalization of key merozoite proteins. AP-2μ is thus essential for blood-stage replication. We generated transgenic P. falciparum parasites expressing hemagglutinin-tagged AP-2μ and examined cellular localization by fluorescence and electron microscopy. Together with mass spectrometry analysis of coimmunoprecipitating proteins, these studies identified AP-2μ-interacting partners, including other AP-2 subunits, the K10 kelch-domain protein, and PfEHD, an effector of endocytosis and lipid mobilization, but no evidence was found of interaction with clathrin, the expected coat protein for AP-2 vesicles. In reverse immunoprecipitation experiments with a clathrin nanobody, other heterotetrameric AP-complexes were shown to interact with clathrin, but AP-2 complex subunits were absent. IMPORTANCE We examine in detail the AP-2 adaptin complex from the malaria parasite Plasmodium falciparum. In most studied organisms, AP-2 is involved in bringing material into the cell from outside, a process called endocytosis. Previous work shows that changes to the μ subunit of AP-2 can contribute to drug resistance. Our experiments show that AP-2 is essential for parasite development in blood but does not have any role in clathrin-mediated endocytosis. This suggests that a specialized function for AP-2 has developed in malaria parasites, and this may be important for understanding its impact on drug resistance. |
| first_indexed | 2024-12-18T01:36:31Z |
| format | Article |
| id | doaj.art-783ffd262b134205b1f7de614ed72d61 |
| institution | Directory Open Access Journal |
| issn | 2150-7511 |
| language | English |
| last_indexed | 2024-12-18T01:36:31Z |
| publishDate | 2020-02-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | mBio |
| spelling | doaj.art-783ffd262b134205b1f7de614ed72d612022-12-21T21:25:26ZengAmerican Society for MicrobiologymBio2150-75112020-02-0111110.1128/mBio.02918-19The <italic toggle="yes">Plasmodium falciparum</italic> Artemisinin Susceptibility-Associated AP-2 Adaptin μ Subunit is Clathrin Independent and Essential for Schizont MaturationRyan C. Henrici0Rachel L. Edwards1Martin Zoltner2Donelly A. van Schalkwyk3Melissa N. Hart4Franziska Mohring5Robert W. Moon6Stephanie D. Nofal7Avnish Patel8Christian Flueck9David A. Baker10Audrey R. Odom John11Mark C. Field12Colin J. Sutherland13Department of Infection Biology, Faculty of Infectious Diseases, London School of Hygiene and Tropical Medicine, London, United KingdomDepartment of Pediatrics, Washington University School of Medicine, St. Louis, Missouri, USASchool of Life Sciences, University of Dundee, Dundee, United KingdomDepartment of Infection Biology, Faculty of Infectious Diseases, London School of Hygiene and Tropical Medicine, London, United KingdomDepartment of Infection Biology, Faculty of Infectious Diseases, London School of Hygiene and Tropical Medicine, London, United KingdomDepartment of Infection Biology, Faculty of Infectious Diseases, London School of Hygiene and Tropical Medicine, London, United KingdomDepartment of Infection Biology, Faculty of Infectious Diseases, London School of Hygiene and Tropical Medicine, London, United KingdomDepartment of Pathogen Molecular Biology, Faculty of Infectious Diseases, London School of Hygiene and Tropical Medicine, London, United KingdomDepartment of Pathogen Molecular Biology, Faculty of Infectious Diseases, London School of Hygiene and Tropical Medicine, London, United KingdomDepartment of Pathogen Molecular Biology, Faculty of Infectious Diseases, London School of Hygiene and Tropical Medicine, London, United KingdomDepartment of Pathogen Molecular Biology, Faculty of Infectious Diseases, London School of Hygiene and Tropical Medicine, London, United KingdomDepartment of Pediatrics, Washington University School of Medicine, St. Louis, Missouri, USASchool of Life Sciences, University of Dundee, Dundee, United KingdomDepartment of Infection Biology, Faculty of Infectious Diseases, London School of Hygiene and Tropical Medicine, London, United KingdomABSTRACT The efficacy of current antimalarial drugs is threatened by reduced susceptibility of Plasmodium falciparum to artemisinin, associated with mutations in pfkelch13. Another gene with variants known to modulate the response to artemisinin encodes the μ subunit of the AP-2 adaptin trafficking complex. To elucidate the cellular role of AP-2μ in P. falciparum, we performed a conditional gene knockout, which severely disrupted schizont organization and maturation, leading to mislocalization of key merozoite proteins. AP-2μ is thus essential for blood-stage replication. We generated transgenic P. falciparum parasites expressing hemagglutinin-tagged AP-2μ and examined cellular localization by fluorescence and electron microscopy. Together with mass spectrometry analysis of coimmunoprecipitating proteins, these studies identified AP-2μ-interacting partners, including other AP-2 subunits, the K10 kelch-domain protein, and PfEHD, an effector of endocytosis and lipid mobilization, but no evidence was found of interaction with clathrin, the expected coat protein for AP-2 vesicles. In reverse immunoprecipitation experiments with a clathrin nanobody, other heterotetrameric AP-complexes were shown to interact with clathrin, but AP-2 complex subunits were absent. IMPORTANCE We examine in detail the AP-2 adaptin complex from the malaria parasite Plasmodium falciparum. In most studied organisms, AP-2 is involved in bringing material into the cell from outside, a process called endocytosis. Previous work shows that changes to the μ subunit of AP-2 can contribute to drug resistance. Our experiments show that AP-2 is essential for parasite development in blood but does not have any role in clathrin-mediated endocytosis. This suggests that a specialized function for AP-2 has developed in malaria parasites, and this may be important for understanding its impact on drug resistance.https://journals.asm.org/doi/10.1128/mBio.02918-19Plasmodium falciparumadaptin trafficking complexartemisinin susceptibilityadaptor proteinsendocytosismalaria |
| spellingShingle | Ryan C. Henrici Rachel L. Edwards Martin Zoltner Donelly A. van Schalkwyk Melissa N. Hart Franziska Mohring Robert W. Moon Stephanie D. Nofal Avnish Patel Christian Flueck David A. Baker Audrey R. Odom John Mark C. Field Colin J. Sutherland The <italic toggle="yes">Plasmodium falciparum</italic> Artemisinin Susceptibility-Associated AP-2 Adaptin μ Subunit is Clathrin Independent and Essential for Schizont Maturation mBio Plasmodium falciparum adaptin trafficking complex artemisinin susceptibility adaptor proteins endocytosis malaria |
| title | The <italic toggle="yes">Plasmodium falciparum</italic> Artemisinin Susceptibility-Associated AP-2 Adaptin μ Subunit is Clathrin Independent and Essential for Schizont Maturation |
| title_full | The <italic toggle="yes">Plasmodium falciparum</italic> Artemisinin Susceptibility-Associated AP-2 Adaptin μ Subunit is Clathrin Independent and Essential for Schizont Maturation |
| title_fullStr | The <italic toggle="yes">Plasmodium falciparum</italic> Artemisinin Susceptibility-Associated AP-2 Adaptin μ Subunit is Clathrin Independent and Essential for Schizont Maturation |
| title_full_unstemmed | The <italic toggle="yes">Plasmodium falciparum</italic> Artemisinin Susceptibility-Associated AP-2 Adaptin μ Subunit is Clathrin Independent and Essential for Schizont Maturation |
| title_short | The <italic toggle="yes">Plasmodium falciparum</italic> Artemisinin Susceptibility-Associated AP-2 Adaptin μ Subunit is Clathrin Independent and Essential for Schizont Maturation |
| title_sort | italic toggle yes plasmodium falciparum italic artemisinin susceptibility associated ap 2 adaptin μ subunit is clathrin independent and essential for schizont maturation |
| topic | Plasmodium falciparum adaptin trafficking complex artemisinin susceptibility adaptor proteins endocytosis malaria |
| url | https://journals.asm.org/doi/10.1128/mBio.02918-19 |
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