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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...

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Autores principales: Kühn, Uwe, Buschmann, Juliane, Wahle, Elmar
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cold Spring Harbor Laboratory Press 2017
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.
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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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