Saccharomyces cerevisiae Pta1 is a component of the cleavage/polyadenylation factor (CPF) 3'-end processing complex and functions in pre-mRNA cleavage, poly(A) addition, and transcription termination. In this study, we investigated the role of the N-terminal region of Pta1 in transcription and processing. We report that a deletion of the first 75 amino acids (pta1-Delta75) causes thermosensitive growth, while the deletion of an additional 25 amino acids is lethal. The pta1-Delta75 mutant is defective for snoRNA termination, RNA polymerase II C-terminal domain Ser5-P dephosphorylation, and gene looping but is fully functional for mRNA 3'-end processing. Furthermore, different regions of Pta1 interact with the CPF subunits Ssu72, Pti1, and Ysh1, supporting the idea that Pta1 acts as a scaffold to organize CPF. The first 300 amino acids of Pta1 are sufficient for interactions with Ssu72, which is needed for pre-mRNA cleavage. By the degron-mediated depletion of Pta1, we show that the removal of this essential region leads to a loss of Ssu72, yet surprisingly, in vitro cleavage and polyadenylation remain efficient. In addition, a fragment containing amino acids 1 to 300 suppresses 3'-end processing in wild-type extracts. These findings suggest that the amino terminus of Pta1 has an inhibitory effect and that this effect can be neutralized through the interaction with Ssu72.
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| Gene | Phenotype | Experiment Type | Mutant Information | Strain Background | Chemical | Details |
|---|---|---|---|---|---|---|
| PTA1 | heat sensitivity: increased | classical genetics | conditional Allele: pta1-Δ75 N-terminal truncation, 1-75 | Other | Temperature: elevated temperature, 39 °C | |
| PTA1 | inviable | classical genetics | null Allele: pta1-Δ100 N-terminal truncation, 1-100 | Other | ||
| PTA1 | protein/peptide accumulation: decreased Reporter: Ssu72p | classical genetics | conditional Allele: pta1-td | Other | Temperature: elevated temperature, 37 °C | |
| PTA1 | protein/peptide modification: increased Reporter: Rpo21p | classical genetics | conditional Allele: pta1-Δ75 N-terminal truncation, 1-75 | Other | Temperature: elevated temperature, 39 °C Details: increased phosphorylation of carboxyl terminal domain (CTD) Ser5 | |
| SSU72 | protein/peptide modification: increased Reporter: Rpo21p | classical genetics | conditional Allele: ssu72-td N-degron allele | Other | Temperature: elevated temperature, 37 °C Details: increased phosphorylation of carboxyl terminal domain (CTD) Ser5 |
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 | |
|---|---|---|---|---|---|---|
| PTA1 | CLP1 | Reconstituted Complex | manually curated | Bait-Hit | No Modification | |
| PTA1 | PCF11 | Reconstituted Complex | manually curated | Bait-Hit | No Modification | |
| PTA1 | CFT2 | Reconstituted Complex | manually curated | Bait-Hit | No Modification | |
| PTA1 | FIP1 | Reconstituted Complex | manually curated | Bait-Hit | No Modification | |
| PTA1 | YTH1 | Reconstituted Complex | manually curated | Bait-Hit | No Modification | |
| PTA1 | CFT1 | Reconstituted Complex | manually curated | Bait-Hit | No Modification | |
| PTA1 | PFS2 | Reconstituted Complex | manually curated | Bait-Hit | No Modification | |
| PTA1 | PAP1 | Reconstituted Complex | manually curated | Bait-Hit | No Modification | |
| PTA1 | SSU72 | Two-hybrid | manually curated | Bait-Hit | No Modification | |
| PTA1 | YSH1 | Two-hybrid | manually curated | Bait-Hit | No Modification |