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Consecutive interactions with HSP90 and eEF1A underlie a functional maturation and storage pathway of AID in the cytoplasm

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...

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Autores principales: Methot, Stephen P., Litzler, Ludivine C., Trajtenberg, Felipe, Zahn, Astrid, Robert, Francis, Pelletier, Jerry, Buschiazzo, Alejandro, Magor, Brad G., Di Noia, Javier M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Rockefeller University Press 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4387293/
https://www.ncbi.nlm.nih.gov/pubmed/25824822
http://dx.doi.org/10.1084/jem.20141157
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author Methot, Stephen P.
Litzler, Ludivine C.
Trajtenberg, Felipe
Zahn, Astrid
Robert, Francis
Pelletier, Jerry
Buschiazzo, Alejandro
Magor, Brad G.
Di Noia, Javier M.
author_facet Methot, Stephen P.
Litzler, Ludivine C.
Trajtenberg, Felipe
Zahn, Astrid
Robert, Francis
Pelletier, Jerry
Buschiazzo, Alejandro
Magor, Brad G.
Di Noia, Javier M.
author_sort Methot, Stephen P.
collection PubMed
description 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.
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spelling pubmed-43872932015-10-06 Consecutive interactions with HSP90 and eEF1A underlie a functional maturation and storage pathway of AID in the cytoplasm Methot, Stephen P. Litzler, Ludivine C. Trajtenberg, Felipe Zahn, Astrid Robert, Francis Pelletier, Jerry Buschiazzo, Alejandro Magor, Brad G. Di Noia, Javier M. J Exp Med Article 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. The Rockefeller University Press 2015-04-06 /pmc/articles/PMC4387293/ /pubmed/25824822 http://dx.doi.org/10.1084/jem.20141157 Text en © 2015 Methot et al. This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).
spellingShingle Article
Methot, Stephen P.
Litzler, Ludivine C.
Trajtenberg, Felipe
Zahn, Astrid
Robert, Francis
Pelletier, Jerry
Buschiazzo, Alejandro
Magor, Brad G.
Di Noia, Javier M.
Consecutive interactions with HSP90 and eEF1A underlie a functional maturation and storage pathway of AID in the cytoplasm
title Consecutive interactions with HSP90 and eEF1A underlie a functional maturation and storage pathway of AID in the cytoplasm
title_full Consecutive interactions with HSP90 and eEF1A underlie a functional maturation and storage pathway of AID in the cytoplasm
title_fullStr Consecutive interactions with HSP90 and eEF1A underlie a functional maturation and storage pathway of AID in the cytoplasm
title_full_unstemmed Consecutive interactions with HSP90 and eEF1A underlie a functional maturation and storage pathway of AID in the cytoplasm
title_short Consecutive interactions with HSP90 and eEF1A underlie a functional maturation and storage pathway of AID in the cytoplasm
title_sort consecutive interactions with hsp90 and eef1a underlie a functional maturation and storage pathway of aid in the cytoplasm
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4387293/
https://www.ncbi.nlm.nih.gov/pubmed/25824822
http://dx.doi.org/10.1084/jem.20141157
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