To investigate the role of Pcf11 in transcriptional termination of ncRNA, we initially performed genome-wide analysis to establish generality. However, we elected to further analyze this data set to specifically look for increased Pol II signal in downstream regions of NRD terminators relative to wild type see the Materials and Methods; Supplemental Fig. A general view of pathway choice for Pol II transcription termination is that termination factor assembly depends on Rpb1 C-terminal domain CTD phosphorylation Gudipati et al. Large-scale Survey conditional colony sectoring: Notably for pcf , no termination defects were evident for these three gene classes Fig. PCF11 encodes a third protein component of yeast cleavage and polyadenylation factor I. Grzechnik P , Kufel J.


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Nrd1 binding stimulates Pcf11 recruitment, which in turn releases Nrd1. The nab mutant strain showed only a minor increase of RNA: Similarly, loss of Rrp6 did not reduce the growth rate of clp Supplemental Fig.


ppcf11 Strains used in this work are listed in the Supplemental Material. Transcriptome surveillance by selective termination of noncoding RNA synthesis. Previous Section Next Section. Furthermore, previous studies have indicated that these different domains may have differential importance in PAS-dependent as compared with NRD transcription termination Birse et al. Diagrams, ChIP procedure, and standardization are as described for Figure 4.


Strain Alignment Variant Viewer. To show that the observed RNA: S8BSer2-P marks were again strongly reduced in pcf Fig. Chemical-genomic dissection of the CTD pcf111. Elevated temperature increases the Pol II transcription rate Miguel et al.


NRD1 strain grown in galactose Fig.

Pcf11 orchestrates transcription termination pathways in yeast

NRD may also regulate hundreds of protein-coding genes through the promotion of premature transcription termination Steinmetz et al. However, we elected to further analyze this data set to specifically look for increased Pol II signal in downstream regions of NRD terminators relative to wild type see the Materials and Methods; Supplemental Fig. Yeast Sen1 helicase protects the genome from transcription-associated instability.

Unexpectedly, the Pcf11 CID mutation genetically interacts with the nuclear exosome cofactor exonuclease Rrp6. Click “Interaction Details” to view all interaction annotations and evidence for this locus, including an interaction visualization. Consistently, thiolutin-dependent pcff11 shutoff in pcf was followed by an initially strong increase of ncRNA NELc and its subsequent delayed degradation.


To confirm that the observed Pol II accumulation downstream from the NBS in sen and, by extension, pcf is due to slow progression of polymerase across the downstream gene, we inserted the inducible GAL1 promoter in place of the SNR13 promoter and analyzed transcription kinetics over the GAL Under such conditions, Sen1 may not be efficiently engaged over the proximal regions of NRD terminators and require higher Pcf11 concentration on chromatin to terminate transcription.

Since Rrp6 rapidly degrades NELc in wild type, we compared its accumulation in pcf with a control strain, sen Notably, we did not detect any change in hybrid accumulation between ppcf11 type and sen Fig. Genome-wide analysis shows that most ncRNA are affected by Pcf11 inactivation, indicating the joint termination requirement for Nrd1 and Pcf Primary 43 Additional 42 Reviews Cells were incubated at the nonpermissive temperature for 30 min prior to thiolutin-specific transcription inhibition.


Pcf1 Venn diagram presenting the overlap between Pcf and Nrd1-dependent transcription units.

Pcf11 orchestrates transcription termination pathways in yeast

This argues that the initial Nrd1 recruitment is independent of Pcf11 levels. Pcd11 above the graphs depict loci and the locations of the amplicons. We therefore investigated whether disturbance of Nrd1—Pcf11 exchange on chromatin, as we observed in pcfcorrelates with Pol II CTD phosphorylation.

However, in the regions downstream, Pol II signals strongly accumulated lcf11 both mutants on all three genes. The number of putative Regulators genes that regulate it and Targets genes it regulates for the given locus, based on experimental evidence.

Thiolutin inhibits transcription initiation but has less effect on elongation Tipper DNA hybrids Mischo et al.