Please use this identifier to cite or link to this item:
http://dx.doi.org/10.25673/85797
Title: | Reconstitution of 3′ end processing of mammalian pre-mRNA reveals a central role of RBBP6 |
Author(s): | Schmidt, Moritz Kluge, Florian Sandmeir, Felix Kühn, Uwe Schäfer, Peter Tüting, Christian Ihling, Christian ![]() Conti, Elena Wahle, Elmar |
Issue Date: | 2022 |
Type: | Article |
Language: | English |
Abstract: | The 3′ ends of almost all eukaryotic mRNAs are generated in an essential two-step processing reaction: endonucleolytic cleavage of an extended precursor followed by the addition of a poly(A) tail. By reconstituting the reaction from overproduced and purified proteins, we provide a minimal list of 14 polypeptides that are essential and two that are stimulatory for RNA processing. In a reaction depending on the polyadenylation signal AAUAAA, the reconstituted system cleaves pre-mRNA at a single preferred site corresponding to the one used in vivo. Among the proteins, cleavage factor I stimulates cleavage but is not essential, consistent with its prominent role in alternative polyadenylation. RBBP6 is required, with structural data showing it to contact and presumably activate the endonuclease CPSF73 through its DWNN domain. The C-terminal domain of RNA polymerase II is dispensable. ATP, but not its hydrolysis, supports RNA cleavage by binding to the hClp1 subunit of cleavage factor II with submicromolar affinity. |
URI: | https://opendata.uni-halle.de//handle/1981185920/87749 http://dx.doi.org/10.25673/85797 |
Open Access: | ![]() |
License: | ![]() |
Sponsor/Funder: | Publikationsfonds MLU |
Journal Title: | Genes & development |
Publisher: | HighWire Press |
Publisher Place: | Stanford, Calif. |
Volume: | 36 |
Original Publication: | 10.1101/gad.349217.121. |
Page Start: | 195 |
Page End: | 209 |
Appears in Collections: | Open Access Publikationen der MLU |
Files in This Item:
File | Description | Size | Format | |
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Genes Dev.-2022-Schmidt-195-209.pdf | 7.21 MB | Adobe PDF | ![]() View/Open |