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A Non-Canonical Initiation Site Is Required for Efficient Translation of the Dendritically Localized Shank1 mRNA

Local protein synthesis in dendrites enables neurons to selectively change the protein complement of individual postsynaptic sites. Though it is generally assumed that this mechanism requires tight translational control of dendritically transported mRNAs, it is unclear how translation of dendritic m...

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Autores principales: Studtmann, Katrin, Ölschläger-Schütt, Janin, Buck, Friedrich, Richter, Dietmar, Sala, Carlo, Bockmann, Jürgen, Kindler, Stefan, Kreienkamp, Hans-Jürgen
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3922875/
https://www.ncbi.nlm.nih.gov/pubmed/24533096
http://dx.doi.org/10.1371/journal.pone.0088518
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author Studtmann, Katrin
Ölschläger-Schütt, Janin
Buck, Friedrich
Richter, Dietmar
Sala, Carlo
Bockmann, Jürgen
Kindler, Stefan
Kreienkamp, Hans-Jürgen
author_facet Studtmann, Katrin
Ölschläger-Schütt, Janin
Buck, Friedrich
Richter, Dietmar
Sala, Carlo
Bockmann, Jürgen
Kindler, Stefan
Kreienkamp, Hans-Jürgen
author_sort Studtmann, Katrin
collection PubMed
description Local protein synthesis in dendrites enables neurons to selectively change the protein complement of individual postsynaptic sites. Though it is generally assumed that this mechanism requires tight translational control of dendritically transported mRNAs, it is unclear how translation of dendritic mRNAs is regulated. We have analyzed here translational control elements of the dendritically localized mRNA coding for the postsynaptic scaffold protein Shank1. In its 5′ region, the human Shank1 mRNA exhibits two alternative translation initiation sites (AUG(+1) and AUG(+214)), three canonical upstream open reading frames (uORFs1-3) and a high GC content. In reporter assays, fragments of the 5′UTR with high GC content inhibit translation, suggesting a contribution of secondary structures. uORF3 is most relevant to translation control as it overlaps with the first in frame start codon (AUG(+1)), directing translation initiation to the second in frame start codon (AUG(+214)). Surprisingly, our analysis points to an additional uORF initiated at a non-canonical ACG start codon. Mutation of this start site leads to an almost complete loss of translation initiation at AUG(+1), demonstrating that this unconventional uORF is required for Shank1 synthesis. Our data identify a novel mechanism whereby initiation at a non-canonical site allows for translation of the main Shank1 ORF despite a highly structured 5′UTR.
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spelling pubmed-39228752014-02-14 A Non-Canonical Initiation Site Is Required for Efficient Translation of the Dendritically Localized Shank1 mRNA Studtmann, Katrin Ölschläger-Schütt, Janin Buck, Friedrich Richter, Dietmar Sala, Carlo Bockmann, Jürgen Kindler, Stefan Kreienkamp, Hans-Jürgen PLoS One Research Article Local protein synthesis in dendrites enables neurons to selectively change the protein complement of individual postsynaptic sites. Though it is generally assumed that this mechanism requires tight translational control of dendritically transported mRNAs, it is unclear how translation of dendritic mRNAs is regulated. We have analyzed here translational control elements of the dendritically localized mRNA coding for the postsynaptic scaffold protein Shank1. In its 5′ region, the human Shank1 mRNA exhibits two alternative translation initiation sites (AUG(+1) and AUG(+214)), three canonical upstream open reading frames (uORFs1-3) and a high GC content. In reporter assays, fragments of the 5′UTR with high GC content inhibit translation, suggesting a contribution of secondary structures. uORF3 is most relevant to translation control as it overlaps with the first in frame start codon (AUG(+1)), directing translation initiation to the second in frame start codon (AUG(+214)). Surprisingly, our analysis points to an additional uORF initiated at a non-canonical ACG start codon. Mutation of this start site leads to an almost complete loss of translation initiation at AUG(+1), demonstrating that this unconventional uORF is required for Shank1 synthesis. Our data identify a novel mechanism whereby initiation at a non-canonical site allows for translation of the main Shank1 ORF despite a highly structured 5′UTR. Public Library of Science 2014-02-12 /pmc/articles/PMC3922875/ /pubmed/24533096 http://dx.doi.org/10.1371/journal.pone.0088518 Text en © 2014 Studtmann et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Studtmann, Katrin
Ölschläger-Schütt, Janin
Buck, Friedrich
Richter, Dietmar
Sala, Carlo
Bockmann, Jürgen
Kindler, Stefan
Kreienkamp, Hans-Jürgen
A Non-Canonical Initiation Site Is Required for Efficient Translation of the Dendritically Localized Shank1 mRNA
title A Non-Canonical Initiation Site Is Required for Efficient Translation of the Dendritically Localized Shank1 mRNA
title_full A Non-Canonical Initiation Site Is Required for Efficient Translation of the Dendritically Localized Shank1 mRNA
title_fullStr A Non-Canonical Initiation Site Is Required for Efficient Translation of the Dendritically Localized Shank1 mRNA
title_full_unstemmed A Non-Canonical Initiation Site Is Required for Efficient Translation of the Dendritically Localized Shank1 mRNA
title_short A Non-Canonical Initiation Site Is Required for Efficient Translation of the Dendritically Localized Shank1 mRNA
title_sort non-canonical initiation site is required for efficient translation of the dendritically localized shank1 mrna
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3922875/
https://www.ncbi.nlm.nih.gov/pubmed/24533096
http://dx.doi.org/10.1371/journal.pone.0088518
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