title: Novel membrane-bound eIF2alpha kinase in the flagellar pocket of Trypanosoma brucei.
abstract: Translational control mediated by phosphorylation of the alpha subunit of the eukaryotic initiation factor 2 (eIF2alpha) is central to stress-induced programs of gene expression. Trypanosomatids, important human pathogens, display differentiation processes elicited by contact with the distinct physiological milieu found in their insect vectors and mammalian hosts, likely representing stress situations. Trypanosoma brucei, the agent of African trypanosomiasis, encodes three potential eIF2alpha kinases (TbeIF2K1 to -K3). We show here that TbeIF2K2 is a transmembrane glycoprotein expressed both in procyclic and in bloodstream forms. The catalytic domain of TbeIF2K2 phosphorylates yeast and mammalian eIF2alpha at Ser51. It also phosphorylates the highly unusual form of eIF2alpha found in trypanosomatids specifically at residue Thr169 that corresponds to Ser51 in other eukaryotes. T. brucei eIF2alpha, however, is not a substrate for GCN2 or PKR in vitro. The putative regulatory domain of TbeIF2K2 does not share any sequence similarity with known eIF2alpha kinases. In both procyclic and bloodstream forms TbeIF2K2 is mainly localized in the membrane of the flagellar pocket, an organelle that is the exclusive site of exo- and endocytosis in these parasites. It can also be detected in endocytic compartments but not in lysosomes, suggesting that it is recycled between endosomes and the flagellar pocket. TbeIF2K2 location suggests a relevance in sensing protein or nutrient transport in T. brucei, an organism that relies heavily on posttranscriptional regulatory mechanisms to control gene expression in different environmental conditions. This is the first membrane-associated eIF2alpha kinase described in unicellular eukaryotes.
authors.affiliation: Departamento de Microbiologia, Imunologia e Parasitologia, Universidade Federal de São Paulo, Rua Botucatu, 862, São Paulo, SP 04023-062, Brazil.
authors.forename: Maria Carolina S
authors.initials: MC
authors.lastname: Moraes
authors.forename: Teresa C L
authors.initials: TC
authors.lastname: Jesus
authors.forename: Nilce N
authors.initials: NN
authors.lastname: Hashimoto
authors.forename: Madhusudan
authors.initials: M
authors.lastname: Dey
authors.forename: Kevin J
authors.initials: KJ
authors.lastname: Schwartz
authors.forename: Viviane S
authors.initials: VS
authors.lastname: Alves
authors.forename: Carla C
authors.initials: CC
authors.lastname: Avila
authors.forename: James D
authors.initials: JD
authors.lastname: Bangs
authors.forename: Thomas E
authors.initials: TE
authors.lastname: Dever
authors.forename: Sergio
authors.initials: S
authors.lastname: Schenkman
authors.forename: Beatriz A
authors.initials: BA
authors.lastname: Castilho
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002462:Cell Membrane / Q000201:enzymology*; D011992:Endosomes / Q000201:enzymology; D015852:Eukaryotic Initiation Factor-2 / Q000378:metabolism; D005407:Flagella / Q000201:enzymology*; D006031:Glycosylation; D006801:Humans; D007425:Intracellular Membranes / Q000201:enzymology; D008018:Life Cycle Stages; D008322:Mammals; D008969:Molecular Sequence Data; D010766:Phosphorylation; D010769:Phosphothreonine / Q000378:metabolism; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D015800:Protozoan Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D014346:Trypanosoma brucei brucei / Q000166:cytology* / Q000201:enzymology* / Q000254:growth & development; D019892:eIF-2 Kinase / Q000737:chemistry / Q000378:metabolism*
chemical_list: D015852:Eukaryotic Initiation Factor-2; D015800:Protozoan Proteins; D010769:Phosphothreonine; D019892:eIF-2 Kinase