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Reconstitution of mammalian cleavage factor II involved in 3′ processing of mRNA precursors

Cleavage factor II (CF II) is a poorly characterized component of the multiprotein complex catalyzing 3′ cleavage and polyadenylation of mammalian mRNA precursors. We have reconstituted CF II as a heterodimer of hPcf11 and hClp1. The heterodimer is active in partially reconstituted cleavage reaction...

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Autores principales: Schäfer, Peter, Tüting, Christian, Schönemann, Lars, Kühn, Uwe, Treiber, Thomas, Treiber, Nora, Ihling, Christian, Graber, Anne, Keller, Walter, Meister, Gunter, Sinz, Andrea, Wahle, Elmar
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cold Spring Harbor Laboratory Press 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6239180/
https://www.ncbi.nlm.nih.gov/pubmed/30139799
http://dx.doi.org/10.1261/rna.068056.118
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author Schäfer, Peter
Tüting, Christian
Schönemann, Lars
Kühn, Uwe
Treiber, Thomas
Treiber, Nora
Ihling, Christian
Graber, Anne
Keller, Walter
Meister, Gunter
Sinz, Andrea
Wahle, Elmar
author_facet Schäfer, Peter
Tüting, Christian
Schönemann, Lars
Kühn, Uwe
Treiber, Thomas
Treiber, Nora
Ihling, Christian
Graber, Anne
Keller, Walter
Meister, Gunter
Sinz, Andrea
Wahle, Elmar
author_sort Schäfer, Peter
collection PubMed
description Cleavage factor II (CF II) is a poorly characterized component of the multiprotein complex catalyzing 3′ cleavage and polyadenylation of mammalian mRNA precursors. We have reconstituted CF II as a heterodimer of hPcf11 and hClp1. The heterodimer is active in partially reconstituted cleavage reactions, whereas hClp1 by itself is not. Pcf11 moderately stimulates the RNA 5′ kinase activity of hClp1; the kinase activity is dispensable for RNA cleavage. CF II binds RNA with nanomolar affinity. Binding is mediated mostly by the two zinc fingers in the C-terminal region of hPcf11. RNA is bound without pronounced sequence-specificity, but extended G-rich sequences appear to be preferred. We discuss the possibility that CF II contributes to the recognition of cleavage/polyadenylation substrates through interaction with G-rich far-downstream sequence elements.
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spelling pubmed-62391802019-12-01 Reconstitution of mammalian cleavage factor II involved in 3′ processing of mRNA precursors Schäfer, Peter Tüting, Christian Schönemann, Lars Kühn, Uwe Treiber, Thomas Treiber, Nora Ihling, Christian Graber, Anne Keller, Walter Meister, Gunter Sinz, Andrea Wahle, Elmar RNA Article Cleavage factor II (CF II) is a poorly characterized component of the multiprotein complex catalyzing 3′ cleavage and polyadenylation of mammalian mRNA precursors. We have reconstituted CF II as a heterodimer of hPcf11 and hClp1. The heterodimer is active in partially reconstituted cleavage reactions, whereas hClp1 by itself is not. Pcf11 moderately stimulates the RNA 5′ kinase activity of hClp1; the kinase activity is dispensable for RNA cleavage. CF II binds RNA with nanomolar affinity. Binding is mediated mostly by the two zinc fingers in the C-terminal region of hPcf11. RNA is bound without pronounced sequence-specificity, but extended G-rich sequences appear to be preferred. We discuss the possibility that CF II contributes to the recognition of cleavage/polyadenylation substrates through interaction with G-rich far-downstream sequence elements. Cold Spring Harbor Laboratory Press 2018-12 /pmc/articles/PMC6239180/ /pubmed/30139799 http://dx.doi.org/10.1261/rna.068056.118 Text en © 2018 Schäfer 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 Article
Schäfer, Peter
Tüting, Christian
Schönemann, Lars
Kühn, Uwe
Treiber, Thomas
Treiber, Nora
Ihling, Christian
Graber, Anne
Keller, Walter
Meister, Gunter
Sinz, Andrea
Wahle, Elmar
Reconstitution of mammalian cleavage factor II involved in 3′ processing of mRNA precursors
title Reconstitution of mammalian cleavage factor II involved in 3′ processing of mRNA precursors
title_full Reconstitution of mammalian cleavage factor II involved in 3′ processing of mRNA precursors
title_fullStr Reconstitution of mammalian cleavage factor II involved in 3′ processing of mRNA precursors
title_full_unstemmed Reconstitution of mammalian cleavage factor II involved in 3′ processing of mRNA precursors
title_short Reconstitution of mammalian cleavage factor II involved in 3′ processing of mRNA precursors
title_sort reconstitution of mammalian cleavage factor ii involved in 3′ processing of mrna precursors
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6239180/
https://www.ncbi.nlm.nih.gov/pubmed/30139799
http://dx.doi.org/10.1261/rna.068056.118
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