Alex Francette - Arndt Lab
Systematic Analysis of Paf1 Complex Contributions to the Transcriptome
Eukaryotic cells must utilize a variety of transcription modulating factors to judiciously decide which genes to transcribe and when. Disruptions of one such elongation factor complex, the evolutionarily conserved Paf1C, have been implicated in oncogenesis and generate subunit-specific phenotypes. However, the relative contributions of these proteins in transcription regulation remain incompletely understood. Here, we attempt to discover and delineate immediate and long-term contributions of the Paf1C to nascent transcription. To this end, we have developed conditional depletion alleles of Paf1C subunits in Saccharomyces cerevisiae to assess their individual contributions to active transcription and to compare with strains null for their respective proteins. Through nascent transcript sequencing we find evidence for an active role of Paf1C subunits in regulating RNA polymerase II processivity at coding genes; an effect that is variably evident in stable deletion strains. We additionally demonstrate that Paf1C is necessary for efficient RNA splicing with splicing defects evident only upon long-term subunit loss. Finally, we show subunit-specific effects of Paf1C on protein-coding gene expression particularly on phosphate and iron regulon genes. Iron regulon transcripts appear stabilized in all Paf1C null strains assessed. This work will provide an extensive analysis of both the immediate and extended roles of all subunits of Paf1C in transcription and chromatin regulation. Our elucidation of Paf1C functions as a globally acting transcription regulator has far reaching impacts on our understanding of both fundamental biology of eukaryotes and human diseases of transcription.
Friday, December 3rd, 2021
A219B Langley Hall
12:00 PM