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Published on: August 4, 2019
Mutations in yeast Pcf11, a conserved protein essential for mRNA 3' end processing and transcription termination,
Joel H Graber1, Derick Hoskinson2, Huiyun Liu2
1Mount Desert Island Biological Laboratory, Bar Harbor, ME 04609, USA.
Abstract:
The Pcf11 protein is an essential subunit of the large complex that cleaves and polyadenylates eukaryotic mRNA precursor. It has also been functionally linked to gene-looping, termination of RNA Polymerase II (Pol II) transcripts, and mRNA export. We have examined a poorly characterized but conserved domain (amino acids 142-225) of the Saccharomyces cerevisiae Pcf11 and found that while it is not needed for mRNA 3' end processing or termination downstream of the poly(A) sites of protein-coding genes, its presence improves the interaction with Pol II and the use of transcription terminators near gene promoters. Analysis of genome-wide Pol II occupancy in cells with Pcf11 missing this region, as well as Pcf11 mutated in the Pol II CTD Interacting Domain, indicates that systematic changes in mRNA expression are mediated primarily at the level of transcription. Global expression analysis also shows that a general stress response, involving both activation and suppression of specific gene sets known to be regulated in response to a wide variety of stresses, is induced in the two pcf11 mutants, even though cells are grown in optimal conditions. The mutants also cause an unbalanced expression of cell wall-related genes that does not activate the Cell Wall Integrity pathway but is associated with strong caffeine sensitivity. Based on these findings, we propose that Pcf11 can modulate the expression level of specific functional groups of genes in ways that do not involve its well-characterized role in mRNA 3' end processing.
Insights
The Pcf11 protein
Area of Science:
- Molecular Biology
- Gene Expression Regulation
- Eukaryotic Transcription
Background:
- Pcf11 protein is crucial for mRNA 3' end processing and polyadenylation in eukaryotes.
- Pcf11 is also implicated in gene-looping, RNA Polymerase II (Pol II) transcript termination, and mRNA export.
- A conserved domain (amino acids 142-225) of Saccharomyces cerevisiae Pcf11 is poorly characterized.
Purpose of the Study:
- To investigate the function of the conserved Pcf11 domain (amino acids 142-225).
- To determine the role of this domain in mRNA processing, transcription termination, and gene expression.
- To understand how Pcf11 modulates gene expression and cellular responses.
Main Methods:
- Analysis of Pcf11 mutants lacking the conserved domain or mutated in the Pol II CTD Interacting Domain.
- Genome-wide Pol II occupancy analysis.
- Global gene expression analysis.
Main Results:
- The Pcf11 domain is not essential for mRNA 3' end processing or downstream termination.
- This domain enhances Pol II interaction and the use of terminators near promoters.
- Mutations lead to transcriptional changes, induce a general stress response, and cause unbalanced cell wall gene expression.
- Mutants exhibit caffeine sensitivity without activating the Cell Wall Integrity pathway.
Conclusions:
- Pcf11 modulates gene expression levels of specific functional groups independently of its role in mRNA 3' end processing.
- The Pcf11 domain influences transcription initiation and termination.
- Pcf11 plays a broader role in regulating gene expression and cellular stress responses than previously understood.
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