The Glc7/PP1 phosphatase triggers Paf1C dissociation from RNA polymerase II to enable transcription termination

The mechanisms that control the dynamic composition of RNAPII elongation complexes govern major transitions in the transcription cycle yet are poorly understood. Here, we show that the transcription elongation factor Spt5 determines elongation complex composition to promote productive elongation and the transition to termination. Using an unbiased genetic screen and genomic approaches in Saccharomyces cerevisiae , we provide evidence that dephosphorylation of the Spt5 C-terminal repeat domain (CTR) by the Glc7/PP1 phosphatase is required to dislodge Paf1C from RNAPII near the cleavage and polyadenylation site (CPS). Mutations in Paf1C or the Spt5 CTR that dissociate Paf1C from RNAPII bypass the requirement for two critical regulatory subunits of Glc7. Depletion of Glc7 causes aberrant retention of Paf1C past the CPS and increased readthrough transcription, which is fully suppressed by Paf1C mutations. We conclude that Paf1C antagonizes termination and Glc7-mediated restructuring of the RNAPII elongation complex is a critical step in transcription termination.