Reconstitution of mammalian Cleavage Factor II involved in 3' processing of mRNA precursors
- Peter Schäfer1,
- Christian Tüting1,
- Lars Schönemann1,
- Uwe Kühn2,
- Thomas Treiber3,
- Nora Treiber3,
- Christian Ihling1,
- Anne Graber4,
- Walter Keller4,
- Gunter Meister3,
- Andrea Sinz1 and
- Elmar Wahle1,5
- 1 Martin Luther University Halle-Wittenberg;
- 2 Martin-Luther-Universität Halle-Wittenberg;
- 3 University of Regensburg;
- 4 Biozentrum, University of Basel
- ↵* Corresponding author; email: ewahle{at}biochemtech.uni-halle.de
Abstract
Cleavage factor II (CF II) is a poorly characterized component of the multi-protein 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.
Keywords
- Received July 9, 2018.
- Accepted August 17, 2018.
- Published by Cold Spring Harbor Laboratory Press for the RNA Society
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