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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Autores principales:	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
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2020
Materias:	Plasmodium falciparum adaptin trafficking complex artemisinin susceptibility adaptor proteins endocytosis malaria Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/783ffd262b134205b1f7de614ed72d61
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spelling	oai:doaj.org-article:783ffd262b134205b1f7de614ed72d612021-11-15T15:56:57ZThe <italic toggle="yes">Plasmodium falciparum</italic> Artemisinin Susceptibility-Associated AP-2 Adaptin μ Subunit is Clathrin Independent and Essential for Schizont Maturation10.1128/mBio.02918-192150-7511https://doaj.org/article/783ffd262b134205b1f7de614ed72d612020-02-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.02918-19https://doaj.org/toc/2150-7511ABSTRACT 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.Ryan C. HenriciRachel L. EdwardsMartin ZoltnerDonelly A. van SchalkwykMelissa N. HartFranziska MohringRobert W. MoonStephanie D. NofalAvnish PatelChristian FlueckDavid A. BakerAudrey R. Odom JohnMark C. FieldColin J. SutherlandAmerican Society for MicrobiologyarticlePlasmodium falciparumadaptin trafficking complexartemisinin susceptibilityadaptor proteinsendocytosismalariaMicrobiologyQR1-502ENmBio, Vol 11, Iss 1 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	Plasmodium falciparum adaptin trafficking complex artemisinin susceptibility adaptor proteins endocytosis malaria Microbiology QR1-502
spellingShingle	Plasmodium falciparum adaptin trafficking complex artemisinin susceptibility adaptor proteins endocytosis malaria Microbiology QR1-502 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
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.
format	article
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
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_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_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_sort	<italic toggle="yes">plasmodium falciparum</italic> artemisinin susceptibility-associated ap-2 adaptin μ subunit is clathrin independent and essential for schizont maturation
publisher	American Society for Microbiology
publishDate	2020
url	https://doaj.org/article/783ffd262b134205b1f7de614ed72d61
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The <italic toggle="yes">Plasmodium falciparum</italic> Artemisinin Susceptibility-Associated AP-2 Adaptin μ Subunit is Clathrin Independent and Essential for Schizont Maturation

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