title: Plasmodium falciparum PfSET7: enzymatic characterization and cellular localization of a novel protein methyltransferase in sporozoite, liver and erythrocytic stage parasites.
abstract: Epigenetic control via reversible histone methylation regulates transcriptional activation throughout the malaria parasite genome, controls the repression of multi-copy virulence gene families and determines sexual stage commitment. Plasmodium falciparum encodes ten predicted SET domain-containing protein methyltransferases, six of which have been shown to be refractory to knock-out in blood stage parasites. We have expressed and purified the first recombinant malaria methyltransferase in sufficient quantities to perform a full enzymatic characterization and reveal the ill-defined PfSET7 is an AdoMet-dependent histone H3 lysine methyltransferase with highest activity towards lysines 4 and 9. Steady-state kinetics of the PfSET7 enzyme are similar to previously characterized histone methyltransferase enzymes from other organisms, however, PfSET7 displays specific protein substrate preference towards nucleosomes with pre-existing histone H3 lysine 14 acetylation. Interestingly, PfSET7 localizes to distinct cytoplasmic foci adjacent to the nucleus in erythrocytic and liver stage parasites, and throughout the cytoplasm in salivary gland sporozoites. Characterized recombinant PfSET7 now allows for target based inhibitor discovery. Specific PfSET7 inhibitors can aid in further investigating the biological role of this specific methyltransferase in transmission, hepatic and blood stage parasites, and may ultimately lead to the development of suitable antimalarial drug candidates against this novel class of essential parasite enzymes.
authors.affiliation: Unité Biologie des Interactions Hôte-Parasite, Département de Parasites et Insectes Vecteurs, Institut Pasteur, Paris 75015, France.|CNRS, ERL 9195, Paris 75015, France.|INSERM, UMR 1201, Paris 75015, France.
authors.forename: Patty B
authors.initials: PB
authors.lastname: Chen
authors.affiliation: Unité Biologie des Interactions Hôte-Parasite, Département de Parasites et Insectes Vecteurs, Institut Pasteur, Paris 75015, France.|CNRS, ERL 9195, Paris 75015, France.|INSERM, UMR 1201, Paris 75015, France.
authors.forename: Shuai
authors.initials: S
authors.lastname: Ding
authors.affiliation: Sorbonne Universités, UPMC Univ Paris 06, INSERM U1135, CNRS ERL 8255, Centre d'Immunologie et des Maladies Infectieuses (CIMI-Paris), 91 Bd de l'hôpital, 75013, Paris, France.
authors.forename: Gigliola
authors.initials: G
authors.lastname: Zanghì
authors.affiliation: Sorbonne Universités, UPMC Univ Paris 06, INSERM U1135, CNRS ERL 8255, Centre d'Immunologie et des Maladies Infectieuses (CIMI-Paris), 91 Bd de l'hôpital, 75013, Paris, France.
authors.forename: Valérie
authors.initials: V
authors.lastname: Soulard
authors.affiliation: Department of Chemical Engineering, Imperial College London, South Kensington Campus, London SW7 2AZ, United Kingdom.
authors.forename: Peter A
authors.initials: PA
authors.lastname: DiMaggio
authors.affiliation: Department of Chemistry, Imperial College London, South Kensington Campus, London SW7 2AZ, United Kingdom.
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: Fuchter
authors.affiliation: Unité Biologie des Interactions Hôte-Parasite, Département de Parasites et Insectes Vecteurs, Institut Pasteur, Paris 75015, France.|CNRS, ERL 9195, Paris 75015, France.|INSERM, UMR 1201, Paris 75015, France.
authors.forename: Salah
authors.initials: S
authors.lastname: Mecheri
authors.affiliation: Sorbonne Universités, UPMC Univ Paris 06, INSERM U1135, CNRS ERL 8255, Centre d'Immunologie et des Maladies Infectieuses (CIMI-Paris), 91 Bd de l'hôpital, 75013, Paris, France.|AP HP, Centre Hospitalo-Universitaire Pitié-Salpêtrière, 75013 Paris, France.
authors.forename: Dominique
authors.initials: D
authors.lastname: Mazier
authors.affiliation: Unité Biologie des Interactions Hôte-Parasite, Département de Parasites et Insectes Vecteurs, Institut Pasteur, Paris 75015, France.|CNRS, ERL 9195, Paris 75015, France.|INSERM, UMR 1201, Paris 75015, France.
authors.forename: Artur
authors.initials: A
authors.lastname: Scherf
authors.affiliation: Unité Biologie des Interactions Hôte-Parasite, Département de Parasites et Insectes Vecteurs, Institut Pasteur, Paris 75015, France.|CNRS, ERL 9195, Paris 75015, France.|INSERM, UMR 1201, Paris 75015, France.
authors.forename: Nicholas A
authors.initials: NA
authors.lastname: Malmquist
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000852:Anopheles / Q000469:parasitology; D016367:Baculoviridae / Q000235:genetics / Q000378:metabolism; D003001:Cloning, Molecular; D044127:Epigenesis, Genetic; D004912:Erythrocytes / Q000469:parasitology / Q000648:ultrastructure; D015870:Gene Expression; D011495:Histone-Lysine N-Methyltransferase / Q000235:genetics / Q000378:metabolism*; D006657:Histones / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D008099:Liver / Q000166:cytology / Q000469:parasitology; D009154:Mutation; D010963:Plasmodium falciparum / Q000201:enzymology* / Q000235:genetics / Q000648:ultrastructure; D015800:Protozoan Proteins / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012469:Salivary Glands / Q000469:parasitology; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D061987:Sf9 Cells; D018411:Spodoptera; D034101:Sporozoites / Q000201:enzymology* / Q000648:ultrastructure; D013379:Substrate Specificity; D053058:Trophozoites / Q000201:enzymology* / Q000648:ultrastructure
chemical_list: D006657:Histones; D015800:Protozoan Proteins; D011994:Recombinant Proteins; D011495:Histone-Lysine N-Methyltransferase