Apicoplast isoprenoid precursor synthesis and the molecular basis of fosmidomycin resistance in <i>Toxoplasma gondii</i>

Nair, Sethu C.; Brooks, Carrie F.; Goodman, Christopher D.; Strurm, Angelika; McFadden, Geoffrey I.; Sundriyal, Sandeep; Anglin, Justin L.; Song, Yuanlin; Moreno, Silvia N.J.; Striepen, Boris

doi:10.1084/jem.20110039

Cited by 145 publications

(159 citation statements)

References 47 publications

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“…The mevalonate and deoxy-D-xylulose-5-phosphate (DOXP) pathways produce isopentyl pyrophosphate, which is the precursor for the biosynthesis of longer isoprenoids. Only the DOXP pathway is expressed by the parasite, and deletion of several genes encoding DOXP enzymes is lethal (17). Toxoplasma expresses a bifunctional farnesyl/geranyl diphosphate synthase (TgFPPS) (18) whose activities are required for isoprenoids to be incorporated into cholesterol, dolichols, or isopentyls.…”

Section: Lipids: We All Need Fatmentioning

confidence: 99%

“…Toxoplasma expresses a bifunctional farnesyl/geranyl diphosphate synthase (TgFPPS) (18) whose activities are required for isoprenoids to be incorporated into cholesterol, dolichols, or isopentyls. Deletion of the TgFPPS gene results in parasites with growth defects in specific types of host cells (e.g., the knockout can grow in human foreskin fibroblasts but cannot grow in macrophages) (17,18). TgFPPS knockouts also cannot survive as extracellular parasites for extended periods of time because of a mitochondrial de- fect that is likely due to a loss of ubiquinone, which is an isoprenylated cofactor of the mitochondrial respiratory chain (18).…”

Section: Lipids: We All Need Fatmentioning

confidence: 99%

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Toxoplasma gondii Development of Its Replicative Niche: in Its Host Cell and Beyond

Blader

Koshy

2014

Eukaryot Cell

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b Intracellular pathogens can replicate efficiently only after they manipulate and modify their host cells to create an environment conducive to replication. While diverse cellular pathways are targeted by different pathogens, metabolism, membrane and cytoskeletal architecture formation, and cell death are the three primary cellular processes that are modified by infections. Toxoplasma gondii is an obligate intracellular protozoan that infects ϳ30% of the world's population and causes severe and lifethreatening disease in developing fetuses, in immune-comprised patients, and in certain otherwise healthy individuals who are primarily found in South America. The high prevalence of Toxoplasma in humans is in large part a result of its ability to modulate these three host cell processes. Here, we highlight recent work defining the mechanisms by which Toxoplasma interacts with these processes. In addition, we hypothesize why some processes are modified not only in the infected host cell but also in neighboring uninfected cells.T oxoplasma gondii is a protozoan, obligate intracellular parasite that is considered one of the world's most successful pathogens (1). Multiple factors contribute to this success, including a complex life cycle in which the parasite can be transmitted by both vertical and horizontal means, efficient propagation within both its primary (felines) and intermediate hosts, extensive mechanisms to evade and disarm host immunity, an ability to form chronic lifelong infections in intermediate hosts, and a wide host tropism in which the parasite can infect most nucleated cells of warm-blooded animals (2). Central to most of these factors is that Toxoplasma has developed the means to replicate efficiently within the hostile intracellular environment of its host cell. In this review, we highlight recent data that have shed light on how parasite growth is achieved by the parasite interacting with its host cell to manipulate host signaling cascades, transcription, cell survival pathways, and membrane transport. In addition, we discuss how parasites interact with neighboring host cells and propose how this may contribute to establishing a permissive microenvironment to improve its overall success. In particular, we focus on those processes that are essential for the growth of all parasite strains and we refer readers to recent reviews that highlight how polymorphic parasite molecules contribute to Toxoplasma virulence (3-5). NUTRIENT ACQUISITIONAs an obligate intracellular parasite that resides within a nonfusogenic vacuole, Toxoplasma must satisfy its nutritional needs by scavenging essential nutrients from its host cell. These nutrients include carbon sources (glucose and glutamine) to fuel its energy demands, specific amino acids, lipids, and other nutrients. Below, we discuss each of these and highlight pathways and processes that are unique to the parasite that could serve as novel drug targets (Fig. 1). GLUCOSE AND GLUTAMINE POWER THE PARASITEToxoplasma expresses a full complement of glycolytic and tr...

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Section: Lipids: We All Need Fatmentioning

confidence: 99%

Section: Lipids: We All Need Fatmentioning

confidence: 99%

Toxoplasma gondii Development of Its Replicative Niche: in Its Host Cell and Beyond

Blader

Koshy

2014

Eukaryot Cell

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“…P. falciparum requires isoprenoid biosynthesis through the MEP pathway during intraerythrocytic development, the stage of parasite growth responsible for the clinical symptoms of malaria. The genetic locus for the first dedicated enzyme of this pathway (deoxyxylulose-5-phosphate reductoisomerase [DXR]) is resistant to genetic disruption in P. falciparum and the related apicomplexan Toxoplasma gondii, and chemical inhibition of the MEP pathway by the small molecule fosmidomycin is lethal to malaria parasites (8)(9)(10). Fosmidomycin treatment of cells inhibits two enzymes of the MEP pathway (DXR and methylerythritol phosphate cytidyltransferase [IspD]), and growth inhibition by fosmidomycin is rescued by supplementation with downstream isoprenoids, such as isopentenyl pyrophosphate and geranylgeraniol (11,12).…”

mentioning

confidence: 99%

Isoprenoid Biosynthesis Inhibition Disrupts Rab5 Localization and Food Vacuolar Integrity in Plasmodium falciparum

et al. 2013

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The antimalarial agent fosmidomycin is a validated inhibitor of the nonmevalonate isoprenoid biosynthesis (methylerythritol 4-phosphate [MEP]) pathway in the malaria parasite, Plasmodium falciparum. Since multiple classes of prenyltransferase inhibitors kill P. falciparum, we hypothesized that protein prenylation was one of the essential functions of this pathway. We found that MEP pathway inhibition with fosmidomycin reduces protein prenylation, confirming that de novo isoprenoid biosynthesis produces the isoprenyl substrates for protein prenylation. One important group of prenylated proteins is small GTPases, such as Rab family members, which mediate cellular vesicular trafficking. We have found that Rab5 proteins dramatically mislocalize upon fosmidomycin treatment, consistent with a loss of protein prenylation. Fosmidomycin treatment caused marked defects in food vacuolar morphology and integrity, consistent with a defect in Rab-mediated vesicular trafficking. These results provide insights to the biological functions of isoprenoids in malaria parasites and may assist the rational selection of secondary agents that will be useful in combination therapy with new isoprenoid biosynthesis inhibitors.

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“…Hydrolysis of the tert-butyl ester group of 12a−c,f and 14d,e with 20% TFA in CH 2 Cl 2 and subsequent EDC- phosphonates 8a−d as before. 35 P-NMR spectra of 7a−f and 8a indicate that these appear as rotameric mixtures, a known 50,51 phenomenon that was further validated by variable-temperature 31 P-NMR studies.…”

Section: Synthesismentioning

confidence: 67%

“…IC 50 values of the most promising compounds (determined as described in the Supporting Information) are reported in Table 1. Again, differences between the various enzymes were apparent.…”

Section: (Scheme 2)mentioning

confidence: 99%

Synthesis and Bioactivity of β-Substituted Fosmidomycin Analogues Targeting 1-Deoxy-d-xylulose-5-phosphate Reductoisomerase

et al. 2015

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ABSTRACT. In the wake of the escalating development of resistance to currently available drugs, blocking the vital MEP pathway for isoprenoid biosynthesis in pathogens such as Plasmodia or Mycobacteria offers interesting prospects for inhibiting their growth. Although the natural product retro-hydroxamate fosmidomycin and its homologue FR900098 potently inhibit 1-deoxy-D-xylulose-5-phosphate reductoisomerase (Dxr), a key enzyme in the MEP pathway, their in vivo activity is compromised due to poor absorption and a suboptimal pharmacokinetic profile. In an effort to facilitate cellular uptake, we introduced aryl or aralkyl substituents at the β-position of the hydroxamate analogue of FR900098. While direct addition of a β-aryl moiety resulted in poor inhibition, longer linkers between the carbon backbone and the phenyl ring were generally associated with better binding to the enzymes, as well as activity against P. falciparum.X-ray structures of the parasite Dxr-inhibitor complexes show that the modes of binding of the two classes of compounds are in fact different. When the "shorter" compounds were bound, the active site flap was fully ordered and similar to that observed for the retro-hydroxamates. The "longer" compounds generate a substantially different flap structure, in which a key tryptophan residue is displaced, and the aromatic group of the ligand lies between the tryptophan and the methyl group on the hydroxamate. Although the most promising new Dxr inhibitors lack activity against E. coli and M. smegmatis, they proved to be highly potent inhibitors of Plasmodium falciparum in vitro growth.3

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Apicoplast isoprenoid precursor synthesis and the molecular basis of fosmidomycin resistance in Toxoplasma gondii

Abstract: Expression of a bacterial transporter protein in Toxoplasma gondii results in parasite susceptibility to Formidomycin, a drug targeting isoprenoid precursor synthesis.

Cited by 145 publications

References 47 publications

Toxoplasma gondii Development of Its Replicative Niche: in Its Host Cell and Beyond

Toxoplasma gondii Development of Its Replicative Niche: in Its Host Cell and Beyond

Isoprenoid Biosynthesis Inhibition Disrupts Rab5 Localization and Food Vacuolar Integrity in Plasmodium falciparum

Synthesis and Bioactivity of β-Substituted Fosmidomycin Analogues Targeting 1-Deoxy-d-xylulose-5-phosphate Reductoisomerase

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