The Heme Biosynthesis Pathway Is Essential for Plasmodium falciparum Development in Mosquito Stage but Not in Blood Stages

Ke, Hangjun; Sigala, Paul A.; Miura, Kazutoyo; Morrisey, Joanne M.; Mather, Michael W.; Crowley, Jan R.; Henderson, Jeffrey P.; Goldberg, Daniel E.; Long, Carole A.; Vaidya, Akhil B.

doi:10.1074/jbc.m114.615831

Cited by 138 publications

(166 citation statements)

References 60 publications

(59 reference statements)

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“…Our observations contrast with previous data (25,26). In these studies, classical mercurochrome staining detected only very few oocysts in mosquitoes infected with P. berghei alas Ϫ or fech Ϫ parasites and none in mosquitoes infected with P. falciparum fech Ϫ parasites.…”

Section: Normal Mosquito Colonization By Pbgdcontrasting

confidence: 57%

“…As expected from the vital role of the Plasmodium ETC in the mosquito vector, P. berghei and P. falciparum parasites that contain targeted deletions of ALAS and of FECH, which catalyzes the ultimate step in heme synthesis, displayed a complete arrest in vector stage development (25,26). Phenocopy of impaired cellular respiration by a dysfunctional heme biosynthesis is consistent with the notion that Plasmodium hemoproteins function exclusively in the ETC.…”

supporting

confidence: 62%

“…2A). As inferred from successful deletion of other heme biosynthesis target genes (25,26) and a successful PBGD knockout in P. falciparum (29), both genes were readily deleted by double homologous recombination, confirming their dispensable nature during Plasmodium berghei blood infection in vivo. Next, isogenic pbgd Ϫ and urod Ϫ parasite lines were generated through fluorescence-activated cell sorting (FACS) (33).…”

Section: Resultsmentioning

confidence: 85%

“…Recent reports (25,26) have refuted the longstanding notion that Plasmodium de novo heme biosynthesis is essential for blood stage development (27). Strikingly, heme scavenging from host erythrocytes is important for parasite growth, as shown by refractoriness to P. falciparum growth within ferrochelatase (FECH)-deficient erythrocytes from individuals with erythropoietic protoporphyria (28).…”

mentioning

confidence: 85%

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Distinct Prominent Roles for Enzymes of Plasmodium berghei Heme Biosynthesis in Sporozoite and Liver Stage Maturation

2016

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Malarial parasites have evolved complex regulation of heme supply and disposal to adjust to heme-rich and -deprived host environments. In addition to its own pathway for heme biosynthesis, Plasmodium likely harbors mechanisms for heme scavenging from host erythrocytes. Elaborate compartmentalization of de novo heme synthesis into three subcellular locations, including the vestigial plastid organelle, indicates critical roles in life cycle progression. In this study, we systematically profile the essentiality of heme biosynthesis by targeted gene deletion of enzymes in early steps of this pathway. We show that disruption of endogenous heme biosynthesis leads to a first detectable defect in oocyst maturation and sporogony in the Anopheles vector, whereas blood stage propagation, colonization of mosquito midguts, or initiation of oocyst development occurs indistinguishably from that of wild-type parasites. Although sporozoites are produced by parasites lacking an intact pathway for heme biosynthesis, they are absent from mosquito salivary glands, indicative of a vital role for heme biosynthesis only in sporozoite maturation. Rescue of the first defect in sporogony permitted analysis of potential roles in liver stages. We show that liver stage parasites benefit from but do not strictly depend upon their own aminolevulinic acid synthase and that they can scavenge aminolevulinic acid from the host environment. Together, our experimental genetics analysis of Plasmodium enzymes for heme biosynthesis exemplifies remarkable shifts between the use of endogenous and host resources during life cycle progression. Heme is a ubiquitous iron-porphyrin complex that serves as a cofactor of hemoproteins, such as globins, catalases, and cytochromes (1). The iron component of heme permits the binding of diatomic gases, as well as unique enzymatic redox reactions based on its reduction potential from the oxidized ferric (Fe 3ϩ ) to the reduced ferrous (Fe 2ϩ ) state (1). Thus, heme mediates fundamental biological processes, including oxygen transport, antioxidant responses, and cellular respiration, and is, therefore, indispensable for virtually all living organisms. Exceptions, like the parasitic kinetoplastid flagellate Phytomonas serpens that can survive in the complete absence of heme (2), are rare. As a result of this nearly universal dependence on heme, two complementary strategies for heme acquisition have evolved; heme scavenging and de novo heme biosynthesis.Plasmodium parasites are the causative agents of malaria and have adapted to an intraerythrocytic lifestyle during blood infection, the exclusive pathogenic phase of the parasite life cycle. Erythrocytes make up the largest pool of heme in the human body, as they are rich in hemoglobin. In turn, hemoglobin constitutes the major source of amino acids for developing blood stage parasites, which import and digest almost all host hemoglobin, liberating large amounts of heme (3). Free heme acts as a biological Fenton reagent and can thus damage organic molecules by oxidation...

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Section: Normal Mosquito Colonization By Pbgdcontrasting

confidence: 57%

supporting

confidence: 62%

Section: Resultsmentioning

confidence: 85%

mentioning

confidence: 85%

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Distinct Prominent Roles for Enzymes of Plasmodium berghei Heme Biosynthesis in Sporozoite and Liver Stage Maturation

2016

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“…Evidently this is not for ATP synthesis because inhibition of the ETC has little effect on cellular ATP levels [38][39][40], and deletion of the functional b-subunit of ATP synthase can be tolerated in blood stages of P. berghei [31]. Heme biosynthesis, which is reliant on TCA-derived succinyl-CoA, is dispensable in asexual blood stages [41,42], and thus dissipation of reducing power from TCA activity for this pathway is not required. Perhaps, then, only re-oxidation of ubiquinol, which is necessary to support the function of the ubiquinone-dependent dihydroorotate dehydrogenase (DHODH) for pyrimidine synthesis, requires ETC activity in blood stages.…”

mentioning

confidence: 99%

Apicomplexan Energy Metabolism: Carbon Source Promiscuity and the Quiescence Hyperbole

Jacot

Waller

Soldati‐Favre

et al. 2016

Trends in Parasitology

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Pages 15apicomplexan mitochondrion in energy generation has remained unclear, given the glucosereplete intracellular nature of the environmental niches of these parasites and the apparent reduction of mitochondrial metabolic pathways [4][5][6][7]. For example, apicomplexan mitochondria lack 'type I' NADH dehydrogenase (NDH, complex I of the ETC) and many apparently lack the enzymes and transporters required for fatty acid b-oxidation [8,9]. Furthermore, the pyruvate dehydrogenase complex (PDH) responsible for conversion of pyruvate into acetyl-CoA -an obligate fuel for the TCA cycle -is only present in the apicoplast [10]. Consequently, there was no obvious entry point to allow catalysis of glycolytic pyruvate by the TCA cycle [10][11][12][13][14]. Despite this, there is overwhelming evidence that apicomplexans rely on a functional and canonical TCA cycle (as reviewed [4,5,15]), and apicomplexan genome annotations indicate the presence of all enzymes of the TCA cycle. Only one clade of apicomplexans, Cryptosporidium spp., show evidence of outright loss of the TCA cycle, but these taxa retain only a highly reduced mitochondrion, the mitosome, and have also lost their apicoplast [5,8,16].This review highlights how metabolomics technologies coupled to genetic manipulation have helped in unraveling apicomplexan parasite plasticity in carbon source utilization through different life stages, revealing a complex and sometimes counter-intuitive pattern of metabolic gains, losses, and reassignments. These changes have presumably been tailored to allow these parasites to thrive within their various host niches, but may constitute some much-needed Achilles' heels in the fight against these devastating diseases. Plasmodium falciparum and the Glycolytic DeceitGlycolysis has long appeared to be the main source of ATP and NAD(P)H in the erythrocytic stages of the malaria parasites, [5,[17][18][19][20][21]. Indeed, glucose uptake in P. falciparum-infected red blood cells (RBCs) has been observed to increase 75-100-fold compared to uninfected RBC [22][23][24][25]. Up to 93% of glucose is converted directly to lactate in asexual stages [26][ 4 _ T D $ D I F F ] and is ultimately excreted into the surrounding host cell. Perturbation of glucose uptake is detrimental to parasite growth [27,28], suggesting that glycolysis is an important part of the parasite's strategy for rapid proliferation. In a manner analogous to the Warburg effect observed in highly-proliferative cancer lines and other rapidly-growing cells (e.g., yeast and bloodstream Trypanosoma spp.), Plasmodium (in erythrocytic stages) has opted for fast generation of ATP through substrate-level phosphorylation and secretion of lactate as an end-product (aerobic fermentative glycolysis), as opposed to the 'slow but efficient' mitochondrial oxidative phosphorylation and complete oxidation [29]. Reasons for this dependence on glycolytic fermentation remain unclear -after all, blood stages of Plasmodium certainly have access to oxygen -but it may be a way of avoiding excessiv...

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A suggested vital function for eIF‐5A and dhs genes during murine malaria blood‐stage infection

et al. 2016

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The biological function of the post‐translational modification hypusine in the eukaryotic initiation factor 5A ( EIF ‐5A) in eukaryotes is still not understood. Hypusine is formed by two sequential enzymatic steps at a specific lysine residue in the precursor protein EIF ‐5A. One important biological function of EIF ‐5A which was recently identified is the translation of polyproline‐rich mRNA , suggesting its biological relevance in a variety of biological processes. Hypusinated eIF ‐5A controls the proliferation of cancer cells and inflammatory processes in malaria. It was shown that pharmacological inhibition of the enzymes involved in this pathway, deoxyhypusine synthase ( DHS ) and the deoxyhypusine hydroxylase ( DOHH ), arrested the growth of malaria parasites. Down‐regulation of both the malarial eIF ‐5A and dhs genes by in vitro and in vivo silencing led to decreased transcript levels and protein expression and, in turn, to low parasitemia, confirming a critical role of hypusination in eIF ‐5A for proliferation in Plasmodium . To further investigate whether eIF ‐5A and the activating enzyme DHS are essential for Plasmodium erythrocytic stages, targeted gene disruption was performed in the rodent malaria parasite Plasmodium berghei . Full disruption of both genes was not successful; instead parasites harboring the episome for eIF ‐5A and dhs genes were obtained, suggesting that these genes may perform vital functions during the pathogenic blood cell stage. Next, a knock‐in strategy was pursued for both endogenous genes eIF ‐5A and dhs from P. berghei . The latter resulted in viable recombinant parasites, strengthening the observation that they might be essential for proliferation during asexual development of the malaria parasite.

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The Heme Biosynthesis Pathway Is Essential for Plasmodium falciparum Development in Mosquito Stage but Not in Blood Stages

Cited by 138 publications

References 60 publications

Distinct Prominent Roles for Enzymes of Plasmodium berghei Heme Biosynthesis in Sporozoite and Liver Stage Maturation

Distinct Prominent Roles for Enzymes of Plasmodium berghei Heme Biosynthesis in Sporozoite and Liver Stage Maturation

Apicomplexan Energy Metabolism: Carbon Source Promiscuity and the Quiescence Hyperbole

A suggested vital function for eIF‐5A and dhs genes during murine malaria blood‐stage infection

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