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The nuclear poly(A) binding protein of mammals, but not of fission yeast, participates in mRNA polyadenylation
The nuclear poly(A) binding protein (PABPN1) has been suggested, on the basis of biochemical evidence, to play a role in mRNA polyadenylation by strongly increasing the processivity of poly(A) polymerase. While experiments in metazoans have tended to support such a role, the results were not unequiv...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Cold Spring Harbor Laboratory Press
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5340911/ https://www.ncbi.nlm.nih.gov/pubmed/28096519 http://dx.doi.org/10.1261/rna.057026.116 |
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author | Kühn, Uwe Buschmann, Juliane Wahle, Elmar |
author_facet | Kühn, Uwe Buschmann, Juliane Wahle, Elmar |
author_sort | Kühn, Uwe |
collection | PubMed |
description | The nuclear poly(A) binding protein (PABPN1) has been suggested, on the basis of biochemical evidence, to play a role in mRNA polyadenylation by strongly increasing the processivity of poly(A) polymerase. While experiments in metazoans have tended to support such a role, the results were not unequivocal, and genetic data show that the S. pombe ortholog of PABPN1, Pab2, is not involved in mRNA polyadenylation. The specific model in which PABPN1 increases the rate of poly(A) tail elongation has never been examined in vivo. Here, we have used 4-thiouridine pulse-labeling to examine the lengths of newly synthesized poly(A) tails in human cells. Knockdown of PABPN1 strongly reduced the synthesis of full-length tails of ∼250 nucleotides, as predicted from biochemical data. We have also purified S. pombe Pab2 and the S. pombe poly(A) polymerase, Pla1, and examined their in vitro activities. Whereas PABPN1 strongly increases the activity of its cognate poly(A) polymerase in vitro, Pab2 was unable to stimulate Pla1 to any significant extent. Thus, in vitro and in vivo data are consistent in supporting a role of PABPN1 but not S. pombe Pab2 in the polyadenylation of mRNA precursors. |
format | Online Article Text |
id | pubmed-5340911 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Cold Spring Harbor Laboratory Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-53409112018-04-01 The nuclear poly(A) binding protein of mammals, but not of fission yeast, participates in mRNA polyadenylation Kühn, Uwe Buschmann, Juliane Wahle, Elmar RNA Report The nuclear poly(A) binding protein (PABPN1) has been suggested, on the basis of biochemical evidence, to play a role in mRNA polyadenylation by strongly increasing the processivity of poly(A) polymerase. While experiments in metazoans have tended to support such a role, the results were not unequivocal, and genetic data show that the S. pombe ortholog of PABPN1, Pab2, is not involved in mRNA polyadenylation. The specific model in which PABPN1 increases the rate of poly(A) tail elongation has never been examined in vivo. Here, we have used 4-thiouridine pulse-labeling to examine the lengths of newly synthesized poly(A) tails in human cells. Knockdown of PABPN1 strongly reduced the synthesis of full-length tails of ∼250 nucleotides, as predicted from biochemical data. We have also purified S. pombe Pab2 and the S. pombe poly(A) polymerase, Pla1, and examined their in vitro activities. Whereas PABPN1 strongly increases the activity of its cognate poly(A) polymerase in vitro, Pab2 was unable to stimulate Pla1 to any significant extent. Thus, in vitro and in vivo data are consistent in supporting a role of PABPN1 but not S. pombe Pab2 in the polyadenylation of mRNA precursors. Cold Spring Harbor Laboratory Press 2017-04 /pmc/articles/PMC5340911/ /pubmed/28096519 http://dx.doi.org/10.1261/rna.057026.116 Text en © 2017 Kühn et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society http://creativecommons.org/licenses/by-nc/4.0/ This article is distributed exclusively by the RNA Society for the first 12 months after the full-issue publication date (see http://rnajournal.cshlp.org/site/misc/terms.xhtml). After 12 months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/. |
spellingShingle | Report Kühn, Uwe Buschmann, Juliane Wahle, Elmar The nuclear poly(A) binding protein of mammals, but not of fission yeast, participates in mRNA polyadenylation |
title | The nuclear poly(A) binding protein of mammals, but not of fission yeast, participates in mRNA polyadenylation |
title_full | The nuclear poly(A) binding protein of mammals, but not of fission yeast, participates in mRNA polyadenylation |
title_fullStr | The nuclear poly(A) binding protein of mammals, but not of fission yeast, participates in mRNA polyadenylation |
title_full_unstemmed | The nuclear poly(A) binding protein of mammals, but not of fission yeast, participates in mRNA polyadenylation |
title_short | The nuclear poly(A) binding protein of mammals, but not of fission yeast, participates in mRNA polyadenylation |
title_sort | nuclear poly(a) binding protein of mammals, but not of fission yeast, participates in mrna polyadenylation |
topic | Report |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5340911/ https://www.ncbi.nlm.nih.gov/pubmed/28096519 http://dx.doi.org/10.1261/rna.057026.116 |
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