title: Apicoplast and endoplasmic reticulum cooperate in fatty acid biosynthesis in apicomplexan parasite Toxoplasma gondii.
abstract: Apicomplexan parasites are responsible for high impact human diseases such as malaria, toxoplasmosis, and cryptosporidiosis. These obligate intracellular pathogens are dependent on both de novo lipid biosynthesis as well as the uptake of host lipids for biogenesis of parasite membranes. Genome annotations and biochemical studies indicate that apicomplexan parasites can synthesize fatty acids via a number of different biosynthetic pathways that are differentially compartmentalized. However, the relative contribution of each of these biosynthetic pathways to total fatty acid composition of intracellular parasite stages remains poorly defined. Here, we use a combination of genetic, biochemical, and metabolomic approaches to delineate the contribution of fatty acid biosynthetic pathways in Toxoplasma gondii. Metabolic labeling studies with [(13)C]glucose showed that intracellular tachyzoites synthesized a range of long and very long chain fatty acids (C14:0-26:1). Genetic disruption of the apicoplast-localized type II fatty-acid synthase resulted in greatly reduced synthesis of saturated fatty acids up to 18 carbons long. Ablation of type II fatty-acid synthase activity resulted in reduced intracellular growth that was partially restored by addition of long chain fatty acids. In contrast, synthesis of very long chain fatty acids was primarily dependent on a fatty acid elongation system comprising three elongases, two reductases, and a dehydratase that were localized to the endoplasmic reticulum. The function of these enzymes was confirmed by heterologous expression in yeast. This elongase pathway appears to have a unique role in generating very long unsaturated fatty acids (C26:1) that cannot be salvaged from the host.
authors.affiliation: Department of Cellular Biology, Center for Tropical and Emerging Global Diseases, University of Georgia, Athens,Georgia 30602, USA.
authors.forename: Srinivasan
authors.initials: S
authors.lastname: Ramakrishnan
authors.forename: Melissa D
authors.initials: MD
authors.lastname: Docampo
authors.forename: James I
authors.initials: JI
authors.lastname: Macrae
authors.forename: François M
authors.initials: FM
authors.lastname: Pujol
authors.forename: Carrie F
authors.initials: CF
authors.lastname: Brooks
authors.forename: Giel G
authors.initials: GG
authors.lastname: van Dooren
authors.forename: J Kalervo
authors.initials: JK
authors.lastname: Hiltunen
authors.forename: Alexander J
authors.initials: AJ
authors.lastname: Kastaniotis
authors.forename: Malcolm J
authors.initials: MJ
authors.lastname: McConville
authors.forename: Boris
authors.initials: B
authors.lastname: Striepen
mesh_terms: D004721:Endoplasmic Reticulum / Q000201:enzymology* / Q000235:genetics; D054889:Fatty Acid Synthase, Type II / Q000235:genetics / Q000378:metabolism*; D005231:Fatty Acids, Unsaturated / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D007553:Isotope Labeling; D055432:Metabolomics / Q000379:methods; D015800:Protozoan Proteins / Q000235:genetics / Q000378:metabolism*; D014122:Toxoplasma / Q000201:enzymology* / Q000235:genetics
chemical_list: D005231:Fatty Acids, Unsaturated; D015800:Protozoan Proteins; D054889:Fatty Acid Synthase, Type II