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Efficient Translation of Dnmt1 Requires Cytoplasmic Polyadenylation and Musashi Binding Elements

Regulation of DNMT1 is critical for epigenetic control of many genes and for genome stability. Using phylogenetic analysis we characterized a block of 27 nucleotides in the 3′UTR of Dnmt1 mRNA identical between humans and Xenopus and investigated the role of the individual elements contained within...

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Autores principales: Rutledge, Charlotte E., Lau, Ho-Tak, Mangan, Hazel, Hardy, Linda L., Sunnotel, Olaf, Guo, Fan, MacNicol, Angus M., Walsh, Colum P., Lees-Murdock, Diane J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3930535/
https://www.ncbi.nlm.nih.gov/pubmed/24586322
http://dx.doi.org/10.1371/journal.pone.0088385
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author Rutledge, Charlotte E.
Lau, Ho-Tak
Mangan, Hazel
Hardy, Linda L.
Sunnotel, Olaf
Guo, Fan
MacNicol, Angus M.
Walsh, Colum P.
Lees-Murdock, Diane J.
author_facet Rutledge, Charlotte E.
Lau, Ho-Tak
Mangan, Hazel
Hardy, Linda L.
Sunnotel, Olaf
Guo, Fan
MacNicol, Angus M.
Walsh, Colum P.
Lees-Murdock, Diane J.
author_sort Rutledge, Charlotte E.
collection PubMed
description Regulation of DNMT1 is critical for epigenetic control of many genes and for genome stability. Using phylogenetic analysis we characterized a block of 27 nucleotides in the 3′UTR of Dnmt1 mRNA identical between humans and Xenopus and investigated the role of the individual elements contained within it. This region contains a cytoplasmic polyadenylation element (CPE) and a Musashi binding element (MBE), with CPE binding protein 1 (CPEB1) known to bind to the former in mouse oocytes. The presence of these elements usually indicates translational control by elongation and shortening of the poly(A) tail in the cytoplasm of the oocyte and in some somatic cell types. We demonstrate for the first time cytoplasmic polyadenylation of Dnmt1 during periods of oocyte growth in mouse and during oocyte activation in Xenopus. Furthermore we show by RNA immunoprecipitation that Musashi1 (MSI1) binds to the MBE and that this element is required for polyadenylation in oocytes. As well as a role in oocytes, site-directed mutagenesis and reporter assays confirm that mutation of either the MBE or CPE reduce DNMT1 translation in somatic cells, but likely act in the same pathway: deletion of the whole conserved region has more severe effects on translation in both ES and differentiated cells. In adult cells lacking MSI1 there is a greater dependency on the CPE, with depletion of CPEB1 or CPEB4 by RNAi resulting in substantially reduced levels of endogenous DNMT1 protein and concurrent upregulation of the well characterised CPEB target mRNA cyclin B1. Our findings demonstrate that CPE- and MBE-mediated translation regulate DNMT1 expression, representing a novel mechanism of post-transcriptional control for this gene.
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spelling pubmed-39305352014-02-25 Efficient Translation of Dnmt1 Requires Cytoplasmic Polyadenylation and Musashi Binding Elements Rutledge, Charlotte E. Lau, Ho-Tak Mangan, Hazel Hardy, Linda L. Sunnotel, Olaf Guo, Fan MacNicol, Angus M. Walsh, Colum P. Lees-Murdock, Diane J. PLoS One Research Article Regulation of DNMT1 is critical for epigenetic control of many genes and for genome stability. Using phylogenetic analysis we characterized a block of 27 nucleotides in the 3′UTR of Dnmt1 mRNA identical between humans and Xenopus and investigated the role of the individual elements contained within it. This region contains a cytoplasmic polyadenylation element (CPE) and a Musashi binding element (MBE), with CPE binding protein 1 (CPEB1) known to bind to the former in mouse oocytes. The presence of these elements usually indicates translational control by elongation and shortening of the poly(A) tail in the cytoplasm of the oocyte and in some somatic cell types. We demonstrate for the first time cytoplasmic polyadenylation of Dnmt1 during periods of oocyte growth in mouse and during oocyte activation in Xenopus. Furthermore we show by RNA immunoprecipitation that Musashi1 (MSI1) binds to the MBE and that this element is required for polyadenylation in oocytes. As well as a role in oocytes, site-directed mutagenesis and reporter assays confirm that mutation of either the MBE or CPE reduce DNMT1 translation in somatic cells, but likely act in the same pathway: deletion of the whole conserved region has more severe effects on translation in both ES and differentiated cells. In adult cells lacking MSI1 there is a greater dependency on the CPE, with depletion of CPEB1 or CPEB4 by RNAi resulting in substantially reduced levels of endogenous DNMT1 protein and concurrent upregulation of the well characterised CPEB target mRNA cyclin B1. Our findings demonstrate that CPE- and MBE-mediated translation regulate DNMT1 expression, representing a novel mechanism of post-transcriptional control for this gene. Public Library of Science 2014-02-20 /pmc/articles/PMC3930535/ /pubmed/24586322 http://dx.doi.org/10.1371/journal.pone.0088385 Text en © 2014 Rutledge 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
Rutledge, Charlotte E.
Lau, Ho-Tak
Mangan, Hazel
Hardy, Linda L.
Sunnotel, Olaf
Guo, Fan
MacNicol, Angus M.
Walsh, Colum P.
Lees-Murdock, Diane J.
Efficient Translation of Dnmt1 Requires Cytoplasmic Polyadenylation and Musashi Binding Elements
title Efficient Translation of Dnmt1 Requires Cytoplasmic Polyadenylation and Musashi Binding Elements
title_full Efficient Translation of Dnmt1 Requires Cytoplasmic Polyadenylation and Musashi Binding Elements
title_fullStr Efficient Translation of Dnmt1 Requires Cytoplasmic Polyadenylation and Musashi Binding Elements
title_full_unstemmed Efficient Translation of Dnmt1 Requires Cytoplasmic Polyadenylation and Musashi Binding Elements
title_short Efficient Translation of Dnmt1 Requires Cytoplasmic Polyadenylation and Musashi Binding Elements
title_sort efficient translation of dnmt1 requires cytoplasmic polyadenylation and musashi binding elements
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3930535/
https://www.ncbi.nlm.nih.gov/pubmed/24586322
http://dx.doi.org/10.1371/journal.pone.0088385
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