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RNA-binding proteins distinguish between similar sequence motifs to promote targeted deadenylation by Ccr4-Not
The Ccr4-Not complex removes mRNA poly(A) tails to regulate eukaryotic mRNA stability and translation. RNA-binding proteins contribute to specificity by interacting with both Ccr4-Not and target mRNAs, but this is not fully understood. Here, we reconstitute accelerated and selective deadenylation of...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
eLife Sciences Publications, Ltd
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6340701/ https://www.ncbi.nlm.nih.gov/pubmed/30601114 http://dx.doi.org/10.7554/eLife.40670 |
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author | Webster, Michael W Stowell, James AW Passmore, Lori A |
author_facet | Webster, Michael W Stowell, James AW Passmore, Lori A |
author_sort | Webster, Michael W |
collection | PubMed |
description | The Ccr4-Not complex removes mRNA poly(A) tails to regulate eukaryotic mRNA stability and translation. RNA-binding proteins contribute to specificity by interacting with both Ccr4-Not and target mRNAs, but this is not fully understood. Here, we reconstitute accelerated and selective deadenylation of RNAs containing AU-rich elements (AREs) and Pumilio-response elements (PREs). We find that the fission yeast homologues of Tristetraprolin/TTP and Pumilio/Puf (Zfs1 and Puf3) interact with Ccr4-Not via multiple regions within low-complexity sequences, suggestive of a multipartite interface that extends beyond previously defined interactions. Using a two-color assay to simultaneously monitor poly(A) tail removal from different RNAs, we demonstrate that Puf3 can distinguish between RNAs of very similar sequence. Analysis of binding kinetics reveals that this is primarily due to differences in dissociation rate constants. Consequently, motif quality is a major determinant of mRNA stability for Puf3 targets in vivo and can be used for the prediction of mRNA targets. |
format | Online Article Text |
id | pubmed-6340701 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | eLife Sciences Publications, Ltd |
record_format | MEDLINE/PubMed |
spelling | pubmed-63407012019-01-24 RNA-binding proteins distinguish between similar sequence motifs to promote targeted deadenylation by Ccr4-Not Webster, Michael W Stowell, James AW Passmore, Lori A eLife Biochemistry and Chemical Biology The Ccr4-Not complex removes mRNA poly(A) tails to regulate eukaryotic mRNA stability and translation. RNA-binding proteins contribute to specificity by interacting with both Ccr4-Not and target mRNAs, but this is not fully understood. Here, we reconstitute accelerated and selective deadenylation of RNAs containing AU-rich elements (AREs) and Pumilio-response elements (PREs). We find that the fission yeast homologues of Tristetraprolin/TTP and Pumilio/Puf (Zfs1 and Puf3) interact with Ccr4-Not via multiple regions within low-complexity sequences, suggestive of a multipartite interface that extends beyond previously defined interactions. Using a two-color assay to simultaneously monitor poly(A) tail removal from different RNAs, we demonstrate that Puf3 can distinguish between RNAs of very similar sequence. Analysis of binding kinetics reveals that this is primarily due to differences in dissociation rate constants. Consequently, motif quality is a major determinant of mRNA stability for Puf3 targets in vivo and can be used for the prediction of mRNA targets. eLife Sciences Publications, Ltd 2019-01-02 /pmc/articles/PMC6340701/ /pubmed/30601114 http://dx.doi.org/10.7554/eLife.40670 Text en © 2019, Webster et al http://creativecommons.org/licenses/by/4.0/ http://creativecommons.org/licenses/by/4.0/This article is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited. |
spellingShingle | Biochemistry and Chemical Biology Webster, Michael W Stowell, James AW Passmore, Lori A RNA-binding proteins distinguish between similar sequence motifs to promote targeted deadenylation by Ccr4-Not |
title | RNA-binding proteins distinguish between similar sequence motifs to promote targeted deadenylation by Ccr4-Not |
title_full | RNA-binding proteins distinguish between similar sequence motifs to promote targeted deadenylation by Ccr4-Not |
title_fullStr | RNA-binding proteins distinguish between similar sequence motifs to promote targeted deadenylation by Ccr4-Not |
title_full_unstemmed | RNA-binding proteins distinguish between similar sequence motifs to promote targeted deadenylation by Ccr4-Not |
title_short | RNA-binding proteins distinguish between similar sequence motifs to promote targeted deadenylation by Ccr4-Not |
title_sort | rna-binding proteins distinguish between similar sequence motifs to promote targeted deadenylation by ccr4-not |
topic | Biochemistry and Chemical Biology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6340701/ https://www.ncbi.nlm.nih.gov/pubmed/30601114 http://dx.doi.org/10.7554/eLife.40670 |
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