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Recognition of polyadenylation sites in yeast pre-mRNAs by cleavage and polyadenylation factor

Dichtl, Bernhard and Keller, Walter 2001, Recognition of polyadenylation sites in yeast pre-mRNAs by cleavage and polyadenylation factor, EMBO journal, vol. 20, no. 12, pp. 3197-3209, doi: 10.1093/emboj/20.12.3197.

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Title Recognition of polyadenylation sites in yeast pre-mRNAs by cleavage and polyadenylation factor
Author(s) Dichtl, BernhardORCID iD for Dichtl, Bernhard orcid.org/0000-0001-5514-4982
Keller, Walter
Journal name EMBO journal
Volume number 20
Issue number 12
Start page 3197
End page 3209
Total pages 13
Publisher Nature Publishing Group
Place of publication London, England
Publication date 2001-06-15
ISSN 0261-4189
1460-2075
Keyword(s) mRNA
polyadenylation signals
pre-mRNA 3'-end processing
RNA–protein interactions
Summary Recognition of poly(A) sites in yeast pre-mRNAs is poorly understood. Employing an in vitro cleavage system with cleavage and polyadenylation factor (CPF) and cleavage factor IA we show that the efficiency and positioning elements are dispensable for poly(A)-site recognition within a short CYC1 substrate in vitro. Instead, U-rich elements immediately upstream and downstream of the poly(A) site mediate cleavage-site recognition within CYC1 and ADH1 pre-mRNAs. These elements act in concert with the poly(A) site to produce multiple recognition sites for the processing machinery, since combinations of mutations within these elements were most effective in cleavage inhibition. Intriguingly, introduction of a U-rich element downstream of the GAL7 poly(A) site strongly enhanced cleavage, underscoring the importance of downstream sequences in general. RNA- binding analyses demonstrate that cleavage depends on the recognition of the poly(A)-site region by CPF. Consistent with in vitro results, mutation of sequences upstream and downstream of the poly(A) site affected 3'-end formation in vivo. A model for yeast pre-mRNA cleavage-site recognition outlines an unanticipated high conservation of yeast and mammalian 3'-end processing mechanisms.
Language eng
DOI 10.1093/emboj/20.12.3197
Field of Research 060199 Biochemistry and Cell Biology not elsewhere classified
Socio Economic Objective 970106 Expanding Knowledge in the Biological Sciences
HERDC Research category C1.1 Refereed article in a scholarly journal
Copyright notice ©2001, Nature Publishing Group
Persistent URL http://hdl.handle.net/10536/DRO/DU:30039330

Document type: Journal Article
Collection: School of Life and Environmental Sciences
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