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Sumoylation is Required for the Cytoplasmic Accumulation of a Subset of mRNAs

In order to discover novel proteins that promote the nuclear export of newly synthesized mRNAs in mammalian cells, we carried out a limited RNAi screen for proteins required for the proper cytoplasmic distribution of a model intronless mRNA. From this screen we obtained two hits, Ubc9 (SUMO-conjugat...

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Autores principales: Zhang, Hui, Mahadevan, Kohila, Palazzo, Alexander F.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4276922/
https://www.ncbi.nlm.nih.gov/pubmed/25333844
http://dx.doi.org/10.3390/genes5040982
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author Zhang, Hui
Mahadevan, Kohila
Palazzo, Alexander F.
author_facet Zhang, Hui
Mahadevan, Kohila
Palazzo, Alexander F.
author_sort Zhang, Hui
collection PubMed
description In order to discover novel proteins that promote the nuclear export of newly synthesized mRNAs in mammalian cells, we carried out a limited RNAi screen for proteins required for the proper cytoplasmic distribution of a model intronless mRNA. From this screen we obtained two hits, Ubc9 (SUMO-conjugating E2 enzyme) and GANP (germinal center-associated nuclear protein). Depletion of Ubc9 inhibited the proper cytoplasmic distribution of certain overexpressed intronless mRNAs, while depletion of GANP affected all tested mRNAs. Depletion of Sae1, which is also required for sumoylation, partially inhibited the cytoplasmic distribution of our model mRNA. Interestingly, the block in cytoplasmic accumulation in Ubc9-depleted cells could be overcome if an intron was incorporated into the mRNA. Surprisingly, Ubc9-depleted cells had normal nuclear export of newly synthesized intronless mRNAs, indicating that the observed accumulation of the model mRNA in the nuclei of transfected cells was likely due to some more general perturbation. Indeed, depletion of Ubc9, coupled with the overexpression of the intronless mRNAs, caused the redistribution of the nuclear speckle protein SC35 to cytoplasmic foci. Our results suggest that sumoylation may play a role in the proper assembly of mRNPs and/or the distribution of key RNA binding proteins, and may thus contribute to general protein expression patterns.
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spelling pubmed-42769222014-12-30 Sumoylation is Required for the Cytoplasmic Accumulation of a Subset of mRNAs Zhang, Hui Mahadevan, Kohila Palazzo, Alexander F. Genes (Basel) Article In order to discover novel proteins that promote the nuclear export of newly synthesized mRNAs in mammalian cells, we carried out a limited RNAi screen for proteins required for the proper cytoplasmic distribution of a model intronless mRNA. From this screen we obtained two hits, Ubc9 (SUMO-conjugating E2 enzyme) and GANP (germinal center-associated nuclear protein). Depletion of Ubc9 inhibited the proper cytoplasmic distribution of certain overexpressed intronless mRNAs, while depletion of GANP affected all tested mRNAs. Depletion of Sae1, which is also required for sumoylation, partially inhibited the cytoplasmic distribution of our model mRNA. Interestingly, the block in cytoplasmic accumulation in Ubc9-depleted cells could be overcome if an intron was incorporated into the mRNA. Surprisingly, Ubc9-depleted cells had normal nuclear export of newly synthesized intronless mRNAs, indicating that the observed accumulation of the model mRNA in the nuclei of transfected cells was likely due to some more general perturbation. Indeed, depletion of Ubc9, coupled with the overexpression of the intronless mRNAs, caused the redistribution of the nuclear speckle protein SC35 to cytoplasmic foci. Our results suggest that sumoylation may play a role in the proper assembly of mRNPs and/or the distribution of key RNA binding proteins, and may thus contribute to general protein expression patterns. MDPI 2014-10-20 /pmc/articles/PMC4276922/ /pubmed/25333844 http://dx.doi.org/10.3390/genes5040982 Text en © 2014 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Zhang, Hui
Mahadevan, Kohila
Palazzo, Alexander F.
Sumoylation is Required for the Cytoplasmic Accumulation of a Subset of mRNAs
title Sumoylation is Required for the Cytoplasmic Accumulation of a Subset of mRNAs
title_full Sumoylation is Required for the Cytoplasmic Accumulation of a Subset of mRNAs
title_fullStr Sumoylation is Required for the Cytoplasmic Accumulation of a Subset of mRNAs
title_full_unstemmed Sumoylation is Required for the Cytoplasmic Accumulation of a Subset of mRNAs
title_short Sumoylation is Required for the Cytoplasmic Accumulation of a Subset of mRNAs
title_sort sumoylation is required for the cytoplasmic accumulation of a subset of mrnas
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4276922/
https://www.ncbi.nlm.nih.gov/pubmed/25333844
http://dx.doi.org/10.3390/genes5040982
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