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Structural insights into the assembly and polyA signal recognition mechanism of the human CPSF complex
3’ polyadenylation is a key step in eukaryotic mRNA biogenesis. In mammalian cells, this process is dependent on the recognition of the hexanucleotide AAUAAA motif in the pre-mRNA polyadenylation signal by the cleavage and polyadenylation specificity factor (CPSF) complex. A core CPSF complex compri...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
eLife Sciences Publications, Ltd
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5760199/ https://www.ncbi.nlm.nih.gov/pubmed/29274231 http://dx.doi.org/10.7554/eLife.33111 |
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author | Clerici, Marcello Faini, Marco Aebersold, Ruedi Jinek, Martin |
author_facet | Clerici, Marcello Faini, Marco Aebersold, Ruedi Jinek, Martin |
author_sort | Clerici, Marcello |
collection | PubMed |
description | 3’ polyadenylation is a key step in eukaryotic mRNA biogenesis. In mammalian cells, this process is dependent on the recognition of the hexanucleotide AAUAAA motif in the pre-mRNA polyadenylation signal by the cleavage and polyadenylation specificity factor (CPSF) complex. A core CPSF complex comprising CPSF160, WDR33, CPSF30 and Fip1 is sufficient for AAUAAA motif recognition, yet the molecular interactions underpinning its assembly and mechanism of PAS recognition are not understood. Based on cross-linking-coupled mass spectrometry, crystal structure of the CPSF160-WDR33 subcomplex and biochemical assays, we define the molecular architecture of the core human CPSF complex, identifying specific domains involved in inter-subunit interactions. In addition to zinc finger domains in CPSF30, we identify using quantitative RNA-binding assays an N-terminal lysine/arginine-rich motif in WDR33 as a critical determinant of specific AAUAAA motif recognition. Together, these results shed light on the function of CPSF in mediating PAS-dependent RNA cleavage and polyadenylation. |
format | Online Article Text |
id | pubmed-5760199 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | eLife Sciences Publications, Ltd |
record_format | MEDLINE/PubMed |
spelling | pubmed-57601992018-01-10 Structural insights into the assembly and polyA signal recognition mechanism of the human CPSF complex Clerici, Marcello Faini, Marco Aebersold, Ruedi Jinek, Martin eLife Biochemistry and Chemical Biology 3’ polyadenylation is a key step in eukaryotic mRNA biogenesis. In mammalian cells, this process is dependent on the recognition of the hexanucleotide AAUAAA motif in the pre-mRNA polyadenylation signal by the cleavage and polyadenylation specificity factor (CPSF) complex. A core CPSF complex comprising CPSF160, WDR33, CPSF30 and Fip1 is sufficient for AAUAAA motif recognition, yet the molecular interactions underpinning its assembly and mechanism of PAS recognition are not understood. Based on cross-linking-coupled mass spectrometry, crystal structure of the CPSF160-WDR33 subcomplex and biochemical assays, we define the molecular architecture of the core human CPSF complex, identifying specific domains involved in inter-subunit interactions. In addition to zinc finger domains in CPSF30, we identify using quantitative RNA-binding assays an N-terminal lysine/arginine-rich motif in WDR33 as a critical determinant of specific AAUAAA motif recognition. Together, these results shed light on the function of CPSF in mediating PAS-dependent RNA cleavage and polyadenylation. eLife Sciences Publications, Ltd 2017-12-23 /pmc/articles/PMC5760199/ /pubmed/29274231 http://dx.doi.org/10.7554/eLife.33111 Text en © 2017, Clerici 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 Clerici, Marcello Faini, Marco Aebersold, Ruedi Jinek, Martin Structural insights into the assembly and polyA signal recognition mechanism of the human CPSF complex |
title | Structural insights into the assembly and polyA signal recognition mechanism of the human CPSF complex |
title_full | Structural insights into the assembly and polyA signal recognition mechanism of the human CPSF complex |
title_fullStr | Structural insights into the assembly and polyA signal recognition mechanism of the human CPSF complex |
title_full_unstemmed | Structural insights into the assembly and polyA signal recognition mechanism of the human CPSF complex |
title_short | Structural insights into the assembly and polyA signal recognition mechanism of the human CPSF complex |
title_sort | structural insights into the assembly and polya signal recognition mechanism of the human cpsf complex |
topic | Biochemistry and Chemical Biology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5760199/ https://www.ncbi.nlm.nih.gov/pubmed/29274231 http://dx.doi.org/10.7554/eLife.33111 |
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