Reconstitution of mammalian Cleavage Factor II involved in 3’ processing of mRNA precursors

Schaefer, P.; Tueting, C.; Schoenemann, L.; Kuehn, U.; Treiber, T.; Treiber, N.; Ihling, C.; Graber, A.; Keller, W.; Meister, G.; Sinz, A.; Wahle, E.

doi:10.1101/366336

bioRxiv · 10.1101/366336

Reconstitution of mammalian Cleavage Factor II involved in 3’ processing of mRNA precursors

Schaefer, P.·Tueting, C.·Schoenemann, L.·Kuehn, U.·Treiber, T.·Treiber, N.·Ihling, C.·Graber, A.·Keller, W.·Meister, G.·Sinz, A.·Wahle, E.

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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.

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Schaefer, P., Tueting, C., Schoenemann, L., Kuehn, U., Treiber, T., Treiber, N., Ihling, C., Graber, A., Keller, W., Meister, G., Sinz, A., Wahle, E.. 2018-07-10. Reconstitution of mammalian Cleavage Factor II involved in 3’ processing of mRNA precursors. https://doi.org/10.1101/366336

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