title: Regulation of ATG8 membrane association by ATG4 in the parasitic protist Toxoplasma gondii.
abstract: In the process of autophagy, the Atg8 protein is conjugated, through a ubiquitin-like system, to the lipid phosphatidylethanolamine (PE) to associate with the membrane of forming autophagosomes. There, it plays a crucial role in the genesis of these organelles and in autophagy in general. In most eukaryotes, the cysteine peptidase Atg4 processes the C terminus of cytosolic Atg8 to regulate its association with autophagosomal membranes and also delipidates Atg8 to release this protein from membranes. The parasitic protist Toxoplasma gondii contains a functional, yet apparently reduced, autophagic machinery. T. gondii Atg8 homolog, in addition to a cytosolic and occasionally autophagosomal localization, also localizes to the apicoplast, a nonphotosynthetic plastid bounded by four membranes. Our attempts to interfere with TgATG8 function showed that it appears to be essential for parasite multiplication inside its host cell. This protein also displays a peculiar C terminus that does not seem to necessitate processing prior to membrane association and yet an unusually large Toxoplasma homolog of ATG4 is predicted in the parasite genome. A TgATG4 conditional expression mutant that we have generated is severely affected in growth, and displays significant alterations at the organellar level, noticeably with a fragmentation of the mitochondrial network and a loss of the apicoplast. TgATG4-depleted parasites appear to be defective in the recycling of membrane-bound TgATG8. Overall, our data highlight a role for the TgATG8 conjugation pathway in maintaining the homeostasis of the parasite's organelles and suggest that Toxoplasma has evolved a specialized autophagic machinery with original regulation.
authors.affiliation: UMR 5235 CNRS; Universités de Montpellier 1 et 2; Dynamique des Interactions Membranaires Normales et Pathologiques; Montpellier, France.
authors.forename: Marie A
authors.initials: MA
authors.lastname: Kong-Hap
authors.forename: Annabelle
authors.initials: A
authors.lastname: Mouammine
authors.forename: Wassim
authors.initials: W
authors.lastname: Daher
authors.forename: Laurence
authors.initials: L
authors.lastname: Berry
authors.forename: Maryse
authors.initials: M
authors.lastname: Lebrun
authors.forename: Jean-François
authors.initials: JF
authors.lastname: Dubremetz
authors.forename: Sébastien
authors.initials: S
authors.lastname: Besteiro
mesh_terms: D000818:Animals; D063546:Apicoplasts / Q000378:metabolism / Q000648:ultrastructure; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism*; D055785:Gene Knockdown Techniques; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006706:Homeostasis; D006801:Humans; D008297:Male; D008928:Mitochondria / Q000378:metabolism / Q000648:ultrastructure; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D010271:Parasites / Q000166:cytology* / Q000378:metabolism* / Q000648:ultrastructure; D010447:Peptide Hydrolases / Q000378:metabolism; D011485:Protein Binding; D021381:Protein Transport; D015800:Protozoan Proteins / Q000737:chemistry / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D013379:Substrate Specificity; D014122:Toxoplasma / Q000166:cytology* / Q000254:growth & development / Q000378:metabolism* / Q000648:ultrastructure
chemical_list: D015800:Protozoan Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D010447:Peptide Hydrolases
keywords: ATG4; ATG8; Toxoplasma; apicoplast; autophagy; mitochondrion; peptidase; plastid; protease