In the yeast Saccharomyces cerevisiae, pre-mRNA 3'-end processing requires six factors: cleavage factor IA (CF IA), cleavage factor IB (CF IB), cleavage factor II (CF II), polyadenylation factor I (PF I), poly(A) polymerase (Pap1p) and poly(A)-binding protein I (Pab1p). We report the characterization of Pfs2p, a WD-repeat protein previously identified in a multiprotein complex carrying PF I-Pap1p activity. The 3'-end-processing defects of pfs2 mutant strains and the results of immunodepletion and immunoinactivation experiments indicate an essential function for Pfs2p in cleavage and polyadenylation. With a one-step affinity purification method that exploits protein A-tagged Pfs2p, we showed that this protein is part of a CF II-PF I complex. Pull-down experiments with GST fusion proteins revealed direct interactions of Pfs2p with subunits of CF II-PF I and CF IA. These results show that Pfs2p plays an essential role in 3'-end formation by bridging different processing factors and thereby promoting the assembly of the processing complex.
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Increase the total number of rows showing on this page by using the pull-down located below the table, or use the page scroll at the table's top right to browse through the table's pages; use the arrows to the right of a column header to sort by that column; filter the table using the "Filter" box at the top of the table; click on the small "i" buttons located within a cell for an annotation to view further details about experiment type and any other genes involved in the interaction.
| Evidence ID | Analyze ID | Interactor | Interactor Systematic Name | Interactor | Interactor Systematic Name | Allele | Assay | Annotation | Action | Phenotype | SGA score | P-value | Source | Reference | Note |
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Increase the total number of rows showing on this page by using the pull-down located below the table, or use the page scroll at the table's top right to browse through the table's pages; use the arrows to the right of a column header to sort by that column; filter the table using the "Filter" box at the top of the table; click on the small "i" buttons located within a cell for an annotation to view further details about experiment type and any other genes involved in the interaction.
| Interactor | Interactor | Assay | Annotation | Action | Modification | |
|---|---|---|---|---|---|---|
| CFT1 | PFS2 | Affinity Capture-Western | manually curated | Hit-Bait | No Modification | |
| FIP1 | PFS2 | Reconstituted Complex | manually curated | Hit-Bait | No Modification | |
| FIP1 | YSH1 | Affinity Capture-Western | manually curated | Bait-Hit | No Modification | |
| PAP1 | PFS2 | Affinity Capture-Western | manually curated | Hit-Bait | No Modification | |
| PAP1 | FIP1 | Affinity Capture-Western | manually curated | Hit-Bait | No Modification | |
| PTA1 | PFS2 | Affinity Capture-Western | manually curated | Hit-Bait | No Modification | |
| RNA14 | PFS2 | Reconstituted Complex | manually curated | Hit-Bait | No Modification | |
| YSH1 | PFS2 | Affinity Capture-Western | manually curated | Hit-Bait | No Modification | |
| YSH1 | PFS2 | Reconstituted Complex | manually curated | Hit-Bait | No Modification |