Vital and dispensable roles of <i>Plasmodium</i> multidrug resistance transporters during blood‐ and mosquito‐stage development

Rijpma, Sanna R.; Velden, Maarten van der; Annoura, Takeshi; Matz, Joachim M.; Kenthirapalan, Sanketha; Kooij, Taco W. A.; Matuschewski, Kai; Gemert, Geert-Jan van; Vegte‐Bolmer, Marga van de; Siebelink-Stoter, Rianne; Graumans, Wouter; Ramesar, Jai; Klop, Onny; Rüssel, Frans G. M.; Sauerwein, Robert W.; Janse, Chris J.; Franke-Fayard, Blandine; Koenderink, Jan B.

doi:10.1111/mmi.13373

Cited by 11 publications

(12 citation statements)

References 45 publications

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“…This confirmed our PCR results showing that Pb ANKA_0614600 and Pb ANKA_0942100 are probably essential (Figures S2–S4). Additionally, the candidate apicoplast membrane transporter, Pb ANKA_0401200, was reported to be refractory to deletion by Rijpma et al (). We recovered parasites from only one of three transfections with the Pb ANKA_0401200 KO construct, and although our PCR results indicated that integration had occurred (Figure S1), we set out to validate our positive result by Southern blot (Figures S5–S6).…”

Section: Resultsmentioning

confidence: 99%

A genetic screen in rodent malaria parasites identifies five new apicoplast putative membrane transporters, one of which is essential in human malaria parasites

Sayers

Mollard

Buchanan

et al. 2017

Cellular Microbiology

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The malaria-causing parasite, Plasmodium, contains a unique non-photosynthetic plastid known as the apicoplast. The apicoplast is an essential organelle bound by four membranes. Although membrane transporters are attractive drug targets, only two transporters have been characterised in the malaria parasite apicoplast membranes. We selected 27 candidate apicoplast membrane proteins, 20 of which are annotated as putative membrane transporters, and performed a genetic screen in Plasmodium berghei to determine blood stage essentiality and subcellular localisation.Eight apparently essential blood stage genes were identified, three of which were apicoplastlocalised: PbANKA_0614600 (DMT2), PbANKA_0401200 (ABCB4), and PbANKA_0505500.Nineteen candidates could be deleted at the blood stage, four of which were apicoplast-localised.Interestingly, three apicoplast-localised candidates lack a canonical apicoplast targeting signal but do contain conserved N-terminal tyrosines with likely roles in targeting. An inducible knockdown of an essential apicoplast putative membrane transporter, PfDMT2, was only viable when supplemented with isopentenyl diphosphate. Knockdown of PfDMT2 resulted in loss of the apicoplast, identifying PfDMT2 as a crucial apicoplast putative membrane transporter and a candidate for therapeutic intervention.

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Section: Resultsmentioning

confidence: 99%

A genetic screen in rodent malaria parasites identifies five new apicoplast putative membrane transporters, one of which is essential in human malaria parasites

Sayers

Mollard

Buchanan

et al. 2017

Cellular Microbiology

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“…Indeed, of all nineteen targeted pumps, eleven were shown to be refractory to gene deletion (Table S1). In addition to the four MDRs (MDR1, MDR4, MDR6, and MDR7) 22, these include the putative cation transporting ATPase, ATP4, (S. Kenthirapalan, K. Matuschewski, T.W.A. Kooij, unpublished data), ATP6 45 and the V-type proton ATPase subunit G 46, ABCI3, and four predicted aminophospholipid transporters 23.…”

Section: Resultsmentioning

confidence: 99%

“…Due to more efficient experimental and computational methods and access to the entire in vivo life cycle, the majority of such studies have been performed using murine malaria model species, notably Plasmodium berghei , and have focussed on single MTPs (Table S1). Two of our most recent studies have more than doubled the number of targeted genes, and have generated loss-of-function mutants in both P. falciparum and P. berghei 2223. A systematic study of the MDR family demonstrated that four of seven members fulfil essential functions during blood-stage development, highlighting these as potential drug targets 22.…”

Section: Introductionmentioning

confidence: 99%

“…Two of our most recent studies have more than doubled the number of targeted genes, and have generated loss-of-function mutants in both P. falciparum and P. berghei 2223. A systematic study of the MDR family demonstrated that four of seven members fulfil essential functions during blood-stage development, highlighting these as potential drug targets 22. While targeting 35 orphan MTPs, Kenthirapalan et al produced the largest collection of P. berghei knock-out parasites available to date 23.…”

Section: Introductionmentioning

confidence: 99%

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Phylogenetic profiles of all membrane transport proteins of the malaria parasite highlight new drug targets

Weiner

Kooij

2016

Microb Cell

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In order to combat the on-going malaria epidemic, discovery of new drug targets remains vital. Proteins that are essential to survival and specific to malaria parasites are key candidates. To survive within host cells, the parasites need to acquire nutrients and dispose of waste products across multiple membranes. Additionally, like all eukaryotes, they must redistribute ions and organic molecules between their various internal membrane bound compartments. Membrane transport proteins mediate all of these processes and are considered important mediators of drug resistance as well as drug targets in their own right. Recently, using advanced experimental genetic approaches and streamlined life cycle profiling, we generated a large collection of Plasmodium berghei gene deletion mutants and assigned essential gene functions, highlighting potential targets for prophylactic, therapeutic, and transmission-blocking anti-malarial drugs. Here, we present a comprehensive orthology assignment of all Plasmodium falciparum putative membrane transport proteins and provide a detailed overview of the associated essential gene functions obtained through experimental genetics studies in human and murine model parasites. Furthermore, we discuss the phylogeny of selected potential drug targets identified in our functional screen. We extensively discuss the results in the context of the functional assignments obtained using gene targeting available to date.

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“…These studies have for the most part focused on essentiality and growth rate phenotypes in the asexual blood stage of the parasite and have ranged from those which specifically targeted subsets of transporter genes to the large‐scale knockout screen of 2578 P. berghei genes (Bushell et al ., ). The transporters targeted by the small‐scale screens have included the genes encoding 35 ‘orphan transporters’ (Kenthirapalan et al ., ), ATP‐binding cassette (ABC) type pumps (Rijpma et al ., ), and transporters predicted to localise to the membranes of the apicoplast (Sayers et al ., ).…”

Section: The Plasmodium Transportomementioning

confidence: 99%

The transportome of the malaria parasite

Martin

2019

Biological Reviews

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Membrane transport proteins, also known as transporters, control the movement of ions, nutrients, metabolites, and waste products across the membranes of a cell and are central to its biology. Proteins of this type also serve as drug targets and are key players in the phenomenon of drug resistance. The malaria parasite has a relatively reduced transportome, with only approximately 2.5% of its genes encoding transporters. Even so, assigning functions and physiological roles to these proteins, and ascertaining their contributions to drug action and drug resistance, has been very challenging. This review presents a detailed critique and synthesis of the disruption phenotypes, protein subcellular localisations, protein functions (observed or predicted), and links to antimalarial drug resistance for each of the parasite's transporter genes. The breadth and depth of the gene disruption data are particularly impressive, with at least one phenotype determined in the parasite's asexual blood stage for each transporter gene, and multiple phenotypes available for 76% of the genes. Analysis of the curated data set revealed there to be relatively little redundancy in the Plasmodium transportome; almost two‐thirds of the parasite's transporter genes are essential or required for normal growth in the asexual blood stage of the parasite, and this proportion increased to 78% when the disruption phenotypes available for the other parasite life stages were included in the analysis. These observations, together with the finding that 22% of the transportome is implicated in the parasite's resistance to existing antimalarials and/or drugs within the development pipeline, indicate that transporters are likely to serve, or are already serving, as drug targets. Integration of the different biological and bioinformatic data sets also enabled the selection of candidates for transport processes known to be essential for parasite survival, but for which the underlying proteins have thus far remained undiscovered. These include potential transporters of pantothenate, isoleucine, or isopentenyl diphosphate, as well as putative anion‐selective channels that may serve as the pore component of the parasite's ‘new permeation pathways’. Other novel insights into the parasite's biology included the identification of transporters for the potential development of antimalarial treatments, transmission‐blocking drugs, prophylactics, and genetically attenuated vaccines. The syntheses presented herein set a foundation for elucidating the functions and physiological roles of key members of the Plasmodium transportome and, ultimately, to explore and realise their potential as therapeutic targets.

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Vital and dispensable roles of Plasmodium multidrug resistance transporters during blood‐ and mosquito‐stage development

Cited by 11 publications

References 45 publications

A genetic screen in rodent malaria parasites identifies five new apicoplast putative membrane transporters, one of which is essential in human malaria parasites

A genetic screen in rodent malaria parasites identifies five new apicoplast putative membrane transporters, one of which is essential in human malaria parasites

Phylogenetic profiles of all membrane transport proteins of the malaria parasite highlight new drug targets

The transportome of the malaria parasite

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