Myristoyl-CoA:Protein N-Myristoyltransferase, an Essential Enzyme and Potential Drug Target in Kinetoplastid Parasites

Price, Helen P.; Menon, Malini R.; Panethymitaki, Chrysoula; Goulding, David; McKean, Paul G.; Smith, Deborah F.

doi:10.1074/jbc.m211391200

Cited by 168 publications

(207 citation statements)

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“…As expected, the ablation of TbNMT leads to a growth arrest after around >90hrs of induction and ultimately to the death of the parasites (Fig. 1C) (see Price et al, 2003). During induction samples of digitonin extracts of total cells were collected and analyzed by SDS-PAGE.…”

Section: Pomp39 Is Likely Myristoylated and Palmitoylatedsupporting

confidence: 57%

“…The RNAi construct was generated based on a pLew-100 stem-loop construct (Wirtz et al, 1999) with blasticidin as selection marker using a 433bp-insert (nucleotides 158-591 of Tb927.8.2070). The TbNMT RNAi cell line was described by Price et al, 2003. Transfection of the constructs was done as described (McCulloch et al, 2004).…”

Section: Cell Linesmentioning

confidence: 99%

“…It has previously been shown that N-terminal myristoyl-transferase (TbNMT) is responsible for essentially all protein myristoylation in T. brucei (Price et al, 2003). In order to directly investigate the putative myristoylation of POMP39 we produced an RNAi cell line allowing inducible ablation of TbNMT with simultaneous expression of C-terminally tagged POMP39 (POMP39-myc).…”

Section: Pomp39 Is Likely Myristoylated and Palmitoylatedmentioning

confidence: 99%

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A component of the mitochondrial outer membrane proteome of T. brucei probably contains covalent bound fatty acids

Albisetti

Wiese

Schneider

et al. 2015

Experimental Parasitology

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ABSTRACT:A subclass of eukaryotic proteins is subject to modification with fatty acids, the most common of which are palmitic and myristic acid. Protein acylation allows association with cellular membranes in the absence of transmembrane domains. Here we examine POMP39, a protein previously described to be present in the outer mitochondrial membrane proteome (POMP) of the protozoan parasite Trypanosoma brucei. POMP39 lacks canonical transmembrane domains, but is likely both myristoylated and palmitoylated on its N-terminus. Interestingly, the protein is also dually localized on the surface of the mitochondrion as well as in the flagellum of both insect-stage and the bloodstream form of the parasites. Upon abolishing of global protein acylation or mutation of the myristoylation site POMP39 relocates to the cytosol. RNAimediated ablation of the protein neither causes a growth phenotype in insectstage nor bloodstream form trypanosomes.

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Section: Pomp39 Is Likely Myristoylated and Palmitoylatedsupporting

confidence: 57%

Section: Cell Linesmentioning

confidence: 99%

Section: Pomp39 Is Likely Myristoylated and Palmitoylatedmentioning

confidence: 99%

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A component of the mitochondrial outer membrane proteome of T. brucei probably contains covalent bound fatty acids

Albisetti

Wiese

Schneider

et al. 2015

Experimental Parasitology

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“…NMT has been shown to be essential for survival in the bloodstream form of T. brucei (Price et 112 al. 2003;Price et al 2010), in insect stages of L. major (Price et al 2003) and L. donovani (Brannigan 113 et al 2010) and most recently in the rodent malaria parasite P. berghei (Pino et al 2012). 114…”

mentioning

confidence: 99%

N-Myristoyltransferase as a potential drug target in malaria and leishmaniasis

et al. 2013

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SUMMARY 12 13Infections caused by protozoan parasites are among the most widespread and intractable transmissible 14 diseases affecting the developing world, with malaria and leishmaniasis being most costly in terms of 15 morbidity and mortality. Although new drugs are urgently required against both diseases in the face of 16 ever-rising resistance to frontline therapies, very few candidates passing through development 17 pipelines possess a known and novel mode of action. Set in the context of drugs currently in use and 18 under development, we present the evidence for N-myristoyltransferase (NMT), an enzyme that N-19 terminally lipidates a wide range of specific target proteins through post-translational modification, as 20 a potential drug target in malaria and the leishmaniases. We discuss the limitations of current 21 knowledge regarding the downstream targets of this enzyme in protozoa, and our recent progress 22 towards potent cell-active NMT inhibitors against the most clinically-relevant species of parasite. 23Finally, we outline the next steps required in terms of both tools to understand N-myristoylation in 24 protozoan parasites, and the generation of potential development candidates based on the output of 25 our recently-reported high-throughput screens. 26 INTRODUCTION 27Malaria 28 Malaria is a disease caused by infection of a human host with protozoan parasites of the genus 29Plasmodium, and is a devastating global health issue with approximately 200 million cases and 30 1 million deaths in 2010 alone (Murray et al. 2012). The complex life cycle of malaria parasites 31 spreads across two hosts and five host tissues whilst undergoing at least ten distinct morphological 32 transitions (Sturm et al. 2006; Mackinnon and Marsh 2010). Replication of parasites and subsequent 33 rupture of erythrocytes in the intra-erythrocytic stages are responsible for the clinical symptoms of 34 malaria, and the majority of drugs target these asexual (human-host) stages of the life cycle. Some 35 species of malaria, most notably Plasmodium vivax, can exist in a latent liver hypnozoite form that 36 can cause relapse even after clearance of bloodstream parasites (Derbyshire et al. 2012; Rodrigues et 37 al. 2012). Of the five relevant species of human parasite, the vast majority of deaths occur from P. 38 falciparum infections, which is the typical cause of severe malaria (Claessens et al. 2012). This has 39 led to the majority of drug discovery efforts focussing on P. falciparum, typically at the expense of 40 other species. Although the demand for new P. falciparum drugs is in no doubt, P. vivax is 41 responsible for the majority of worldwide malaria endemicity (Price et al. 2009; WHO 2011). 42However, difficulties culturing the parasite (Udomsangpetch et al. 2007) For the latter half of the 20 th century, antimalarial drug discovery was a success story for natural 47 product-inspired therapies, by far the most widely used of which are chloroquine (Loeb et al. 1946) 48 and artemisinin (Miller and Su 2011). 49Chlo...

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“…En 1990 se reportaron los primeros sistemas de transfección permanente en Leishmania. Los ARNm transcritos a partir del ADN plasmídico contenían el ME unido al gen a expresar en la posición correcta del sitio aceptor del empalme; además, estaban poliadenilados (28,103). La transfección permanente se logra con la introducción de episomas o por integración de ADN al genoma.…”

Section: Transfección Estable En Leishmaniaunclassified

Manipulación genética y el estudio del parásito protozoario Leishmania.

Cortázar

Walker

2004

biomedica

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Durante los últimos 15 años se ha dado paso al entendimiento de muchos aspectos de la genómica funcional de Leishmania gracias a los avances en la metodología de transfección de ADN dentro de la célula de este protozoario, la eliminación y la complementación de genes por medio de recombinación homóloga y las estrategias para la selección de células transfectadas. Estos acercamientos tienen el potencial de brindar información sobre la expresión génica y la función de las proteínas en el contexto del parásito intacto. Dado que el genoma de Leishmania muestra una carencia acentuada de los factores conocidos de iniciación de la transcripción y que la expresión génica está regulada casi completamente a nivel postranscripcional (a través del empalme de los ARNm y los mecanismos que involucran el procesamiento diferencial de la región no traducida 3' del ARNm (3'UTR), la transfección génica representa una herramienta útil para la identificación y el análisis funcional de los genes de interés así como de los mecanismos que dirigen su regulación. El desarrollo de los sistemas de manipulación genética también ha abierto nuevos horizontes para la identificación de genes esenciales involucrados en la virulencia, la supervivencia intracelular y la resistencia a drogas de Leishmania, así como para la validación de proteínas específicas del parásito como nuevos blancos quimio e inmunoterapéuticos. En esta revisión presentamos los avances más recientes en el campo de la manipulación genética en Leishmania, los cuales permiten análisis estructurales, funcionales y de fenotipo, por medio de la eliminación y complementación génica a través de la transfección transitoria o permanente de genes en este parásito.Palabras clave: Leishmania, transfección de ADN, expresión génica, eliminación y complementación génica, genómica funcional. Genetic manipulation and the study of the protozoan parasite LeishmaniaDuring the last 15 years, many aspects of the functional genomics of Leishmania have been revealed due to advances in DNA transfection, gene disruption and complementation through homologous recombination, and efficient strategies for the selection of transfected cells. These strategies have provided information about gene expression and protein function in the context of the intact parasite. The genome of Leishmania shows a marked deficiency of known transcription initiation factors, and gene expression is regulated almost entirely at the posttranscriptional level through trans-splicing of mRNAs and novel control mechanisms involving differential processing of 3' -untranslated regions (3'-UTRs) of mRNAs. Therefore, gene transfection represents a useful tool for the identification and functional analysis of genes of interest as well as the mechanisms that direct their regulation. The development of genetic manipulation systems has provided opportunities for the study of genes involved in virulence, intracellular survival and drug resistance of Leishmania, as well as for the functional validation of specific parasite proteins as new ch...

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Myristoyl-CoA:Protein N-Myristoyltransferase, an Essential Enzyme and Potential Drug Target in Kinetoplastid Parasites

Cited by 168 publications

References 50 publications

A component of the mitochondrial outer membrane proteome of T. brucei probably contains covalent bound fatty acids

A component of the mitochondrial outer membrane proteome of T. brucei probably contains covalent bound fatty acids

N-Myristoyltransferase as a potential drug target in malaria and leishmaniasis

Manipulación genética y el estudio del parásito protozoario Leishmania.

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