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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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Detalles Bibliográficos
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
Descripción
Sumario: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.