title: Consecutive interactions with HSP90 and eEF1A underlie a functional maturation and storage pathway of AID in the cytoplasm.
abstract: Activation-induced deaminase (AID) initiates mutagenic pathways to diversify the antibody genes during immune responses. The access of AID to the nucleus is limited by CRM1-mediated nuclear export and by an uncharacterized mechanism of cytoplasmic retention. Here, we define a conformational motif in AID that dictates its cytoplasmic retention and demonstrate that the translation elongation factor eukaryotic elongation factor 1 α (eEF1A) is necessary for AID cytoplasmic sequestering. The mechanism is independent of protein synthesis but dependent on a tRNA-free form of eEF1A. Inhibiting eEF1A prevents the interaction with AID, which accumulates in the nucleus and increases class switch recombination as well as chromosomal translocation byproducts. Most AID is associated to unspecified cytoplasmic complexes. We find that the interactions of AID with eEF1A and heat-shock protein 90 kD (HSP90) are inversely correlated. Despite both interactions stabilizing AID, the nature of the AID fractions associated with HSP90 or eEF1A are different, defining two complexes that sequentially produce and store functional AID in the cytoplasm. In addition, nuclear export and cytoplasmic retention cooperate to exclude AID from the nucleus but might not be functionally equivalent. Our results elucidate the molecular basis of AID cytoplasmic retention, define its functional relevance and distinguish it from other mechanisms regulating AID.
authors.affiliation: Institut de Recherches Cliniques de Montréal, Montréal, Québec H2W 1R7, Canada Department of Medicine, Department of Biochemistry, The Rosalind and Morris Goodman Cancer Research Center, and Department of Oncology, McGill University, Montréal, Québec H3A 0G4, Canada.
authors.forename: Stephen P
authors.initials: SP
authors.lastname: Methot
authors.affiliation: Institut de Recherches Cliniques de Montréal, Montréal, Québec H2W 1R7, Canada Department of Biochemistry and Department of Medicine, Université de Montréal, Montréal, Québec H3T 1J4, Canada.
authors.forename: Ludivine C
authors.initials: LC
authors.lastname: Litzler
authors.affiliation: Unit of Protein Crystallography, Institut Pasteur de Montevideo, Montevideo 11400, Uruguay.
authors.forename: Felipe
authors.initials: F
authors.lastname: Trajtenberg
authors.affiliation: Institut de Recherches Cliniques de Montréal, Montréal, Québec H2W 1R7, Canada.
authors.forename: Astrid
authors.initials: A
authors.lastname: Zahn
authors.affiliation: Department of Medicine, Department of Biochemistry, The Rosalind and Morris Goodman Cancer Research Center, and Department of Oncology, McGill University, Montréal, Québec H3A 0G4, Canada.
authors.forename: Francis
authors.initials: F
authors.lastname: Robert
authors.affiliation: Department of Medicine, Department of Biochemistry, The Rosalind and Morris Goodman Cancer Research Center, and Department of Oncology, McGill University, Montréal, Québec H3A 0G4, Canada Department of Medicine, Department of Biochemistry, The Rosalind and Morris Goodman Cancer Research Center, and Department of Oncology, McGill University, Montréal, Québec H3A 0G4, Canada Department of Medicine, Department of Biochemistry, The Rosalind and Morris Goodman Cancer Research Center, and Department of Oncology, McGill University, Montréal, Québec H3A 0G4, Canada.
authors.forename: Jerry
authors.initials: J
authors.lastname: Pelletier
authors.affiliation: Unit of Protein Crystallography, Institut Pasteur de Montevideo, Montevideo 11400, Uruguay Department of Structural Biology and Chemistry, Institut Pasteur, 75015 Paris, France.
authors.forename: Alejandro
authors.initials: A
authors.lastname: Buschiazzo
authors.affiliation: Department of Biological Sciences, University of Alberta, Edmonton, Alberta T6G 2E9, Canada.
authors.forename: Brad G
authors.initials: BG
authors.lastname: Magor
authors.affiliation: Institut de Recherches Cliniques de Montréal, Montréal, Québec H2W 1R7, Canada Department of Medicine, Department of Biochemistry, The Rosalind and Morris Goodman Cancer Research Center, and Department of Oncology, McGill University, Montréal, Québec H3A 0G4, Canada Department of Biochemistry and Department of Medicine, Université de Montréal, Montréal, Québec H3T 1J4, Canada Department of Biochemistry and Department of Medicine, Université de Montréal, Montréal, Québec H3T 1J4, Canada javier.di.noia@ircm.qc.ca.
authors.forename: Javier M
authors.initials: JM
authors.lastname: Di Noia
mesh_terms: D021581:Active Transport, Cell Nucleus / Q000235:genetics; D020816:Amino Acid Motifs; D000818:Animals; D002467:Cell Nucleus / Q000235:genetics / Q000378:metabolism; D003564:Cytidine Deaminase / Q000235:genetics / Q000378:metabolism*; D003593:Cytoplasm / Q000235:genetics / Q000378:metabolism*; D057809:HEK293 Cells; D006801:Humans; D051379:Mice; D018345:Mice, Knockout; D046912:Multiprotein Complexes / Q000235:genetics / Q000378:metabolism; D020648:Peptide Elongation Factor 1 / Q000235:genetics / Q000378:metabolism*; D014178:Translocation, Genetic; D000098830:AICDA (Activation-Induced Cytidine Deaminase)
chemical_list: D046912:Multiprotein Complexes; D020648:Peptide Elongation Factor 1; D000098830:AICDA (Activation-Induced Cytidine Deaminase); D003564:Cytidine Deaminase
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