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Translation- and SRP-independent mRNA targeting to the endoplasmic reticulum in the yeast Saccharomyces cerevisiae

mRNAs encoding secreted/membrane proteins (mSMPs) are believed to reach the endoplasmic reticulum (ER) in a translation-dependent manner to confer protein translocation. Evidence exists, however, for translation- and signal recognition particle (SRP)–independent mRNA localization to the ER, suggesti...

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Autores principales: Kraut-Cohen, Judith, Afanasieva, Evgenia, Haim-Vilmovsky, Liora, Slobodin, Boris, Yosef, Ido, Bibi, Eitan, Gerst, Jeffrey E.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The American Society for Cell Biology 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3784381/
https://www.ncbi.nlm.nih.gov/pubmed/23904265
http://dx.doi.org/10.1091/mbc.E13-01-0038
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author Kraut-Cohen, Judith
Afanasieva, Evgenia
Haim-Vilmovsky, Liora
Slobodin, Boris
Yosef, Ido
Bibi, Eitan
Gerst, Jeffrey E.
author_facet Kraut-Cohen, Judith
Afanasieva, Evgenia
Haim-Vilmovsky, Liora
Slobodin, Boris
Yosef, Ido
Bibi, Eitan
Gerst, Jeffrey E.
author_sort Kraut-Cohen, Judith
collection PubMed
description mRNAs encoding secreted/membrane proteins (mSMPs) are believed to reach the endoplasmic reticulum (ER) in a translation-dependent manner to confer protein translocation. Evidence exists, however, for translation- and signal recognition particle (SRP)–independent mRNA localization to the ER, suggesting that there are alternate paths for RNA delivery. We localized endogenously expressed mSMPs in yeast using an aptamer-based RNA-tagging procedure and fluorescence microscopy. Unlike mRNAs encoding polarity and secretion factors that colocalize with cortical ER at the bud tip, mSMPs and mRNAs encoding soluble, nonsecreted, nonpolarized proteins localized mainly to ER peripheral to the nucleus (nER). Synthetic nontranslatable uracil-rich mRNAs were also demonstrated to colocalize with nER in yeast. This mRNA–ER association was verified by subcellular fractionation and reverse transcription-PCR, single-molecule fluorescence in situ hybridization, and was not inhibited upon SRP inactivation. To better understand mSMP targeting, we examined aptamer-tagged USE1, which encodes a tail-anchored membrane protein, and SUC2, which encodes a soluble secreted enzyme. USE1 and SUC2 mRNA targeting was not abolished by the inhibition of translation or removal of elements involved in translational control. Overall we show that mSMP targeting to the ER is both translation- and SRP-independent, and regulated by cis elements contained within the message and trans-acting RNA-binding proteins (e.g., She2, Puf2).
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spelling pubmed-37843812013-12-16 Translation- and SRP-independent mRNA targeting to the endoplasmic reticulum in the yeast Saccharomyces cerevisiae Kraut-Cohen, Judith Afanasieva, Evgenia Haim-Vilmovsky, Liora Slobodin, Boris Yosef, Ido Bibi, Eitan Gerst, Jeffrey E. Mol Biol Cell Articles mRNAs encoding secreted/membrane proteins (mSMPs) are believed to reach the endoplasmic reticulum (ER) in a translation-dependent manner to confer protein translocation. Evidence exists, however, for translation- and signal recognition particle (SRP)–independent mRNA localization to the ER, suggesting that there are alternate paths for RNA delivery. We localized endogenously expressed mSMPs in yeast using an aptamer-based RNA-tagging procedure and fluorescence microscopy. Unlike mRNAs encoding polarity and secretion factors that colocalize with cortical ER at the bud tip, mSMPs and mRNAs encoding soluble, nonsecreted, nonpolarized proteins localized mainly to ER peripheral to the nucleus (nER). Synthetic nontranslatable uracil-rich mRNAs were also demonstrated to colocalize with nER in yeast. This mRNA–ER association was verified by subcellular fractionation and reverse transcription-PCR, single-molecule fluorescence in situ hybridization, and was not inhibited upon SRP inactivation. To better understand mSMP targeting, we examined aptamer-tagged USE1, which encodes a tail-anchored membrane protein, and SUC2, which encodes a soluble secreted enzyme. USE1 and SUC2 mRNA targeting was not abolished by the inhibition of translation or removal of elements involved in translational control. Overall we show that mSMP targeting to the ER is both translation- and SRP-independent, and regulated by cis elements contained within the message and trans-acting RNA-binding proteins (e.g., She2, Puf2). The American Society for Cell Biology 2013-10-01 /pmc/articles/PMC3784381/ /pubmed/23904265 http://dx.doi.org/10.1091/mbc.E13-01-0038 Text en © 2013 Kraut-Cohen et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0). “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society of Cell Biology.
spellingShingle Articles
Kraut-Cohen, Judith
Afanasieva, Evgenia
Haim-Vilmovsky, Liora
Slobodin, Boris
Yosef, Ido
Bibi, Eitan
Gerst, Jeffrey E.
Translation- and SRP-independent mRNA targeting to the endoplasmic reticulum in the yeast Saccharomyces cerevisiae
title Translation- and SRP-independent mRNA targeting to the endoplasmic reticulum in the yeast Saccharomyces cerevisiae
title_full Translation- and SRP-independent mRNA targeting to the endoplasmic reticulum in the yeast Saccharomyces cerevisiae
title_fullStr Translation- and SRP-independent mRNA targeting to the endoplasmic reticulum in the yeast Saccharomyces cerevisiae
title_full_unstemmed Translation- and SRP-independent mRNA targeting to the endoplasmic reticulum in the yeast Saccharomyces cerevisiae
title_short Translation- and SRP-independent mRNA targeting to the endoplasmic reticulum in the yeast Saccharomyces cerevisiae
title_sort translation- and srp-independent mrna targeting to the endoplasmic reticulum in the yeast saccharomyces cerevisiae
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3784381/
https://www.ncbi.nlm.nih.gov/pubmed/23904265
http://dx.doi.org/10.1091/mbc.E13-01-0038
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