title: The eIF3 complex of Leishmania-subunit composition and mode of recruitment to different cap-binding complexes.
abstract: Eukaryotic initiation factor 3 (eIF3) is a multi-protein complex and a key participant in the assembly of the translation initiation machinery. In mammals, eIF3 comprises 13 subunits, most of which are characterized by conserved structural domains. The trypanosomatid eIF3 subunits are poorly conserved. Here, we identify 12 subunits that comprise the Leishmania eIF3 complex (LeishIF3a-l) by combining bioinformatics with affinity purification and mass spectrometry analyses. These results highlight the strong association of LeishIF3 with LeishIF1, LeishIF2 and LeishIF5, suggesting the existence of a multi-factor complex. In trypanosomatids, the translation machinery is tightly regulated in the different life stages of these organisms as part of their adaptation and survival in changing environments. We, therefore, addressed the mechanism by which LeishIF3 is recruited to different mRNA cap-binding complexes. A direct interaction was observed in vitro between the fully assembled LeishIF3 complex and recombinant LeishIF4G3, the canonical scaffolding protein of the cap-binding complex in Leishmania promastigotes. We further highlight a novel interaction between the C-terminus of LeishIF3a and LeishIF4E1, the only cap-binding protein that efficiently binds the cap structure under heat shock conditions, anchoring a complex that is deficient of any MIF4G-based scaffolding subunit.
authors.affiliation: Department of Life Sciences, Ben-Gurion University of the Negev, Beer Sheva, Israel.
authors.forename: Shimi
authors.initials: S
authors.lastname: Meleppattu
authors.affiliation: Department of Life Sciences, Ben-Gurion University of the Negev, Beer Sheva, Israel.
authors.forename: Dikla
authors.initials: D
authors.lastname: Kamus-Elimeleh
authors.affiliation: Department of Life Sciences, Ben-Gurion University of the Negev, Beer Sheva, Israel.
authors.forename: Alexandra
authors.initials: A
authors.lastname: Zinoviev
authors.affiliation: Department of Life Sciences, Ben-Gurion University of the Negev, Beer Sheva, Israel.
authors.forename: Shahar
authors.initials: S
authors.lastname: Cohen-Mor
authors.affiliation: Department of Life Sciences, Ben-Gurion University of the Negev, Beer Sheva, Israel.
authors.forename: Irit
authors.initials: I
authors.lastname: Orr
authors.affiliation: Department of Life Sciences, Ben-Gurion University of the Negev, Beer Sheva, Israel shapiram@bgu.ac.il.
authors.forename: Michal
authors.initials: M
authors.lastname: Shapira
mesh_terms: D019295:Computational Biology; D039621:Eukaryotic Initiation Factor-3 / Q000737:chemistry / Q000378:metabolism*; D039562:Eukaryotic Initiation Factor-4F / Q000378:metabolism*; D007894:Leishmania mexicana / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D010442:Peptide Chain Initiation, Translational; D021122:Protein Subunits / Q000378:metabolism; D015800:Protozoan Proteins / Q000737:chemistry / Q000378:metabolism*
chemical_list: D039621:Eukaryotic Initiation Factor-3; D039562:Eukaryotic Initiation Factor-4F; D021122:Protein Subunits; D015800:Protozoan Proteins