2014
DOI: 10.1016/j.celrep.2014.10.007
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The Evolutionarily Conserved Pol II Flap Loop Contributes to Proper Transcription Termination on Short Yeast Genes

Abstract: SUMMARY Current models of transcription termination factor recruitment to the RNA polymerase II (Pol II) transcription complex rely exclusively on the direct interaction between the termination factor and phosphorylated isoforms of the Pol II C-terminal domain (CTD). Here we report that the Pol II flap loop is needed for physical interaction of Pol II with the Pcf11/Clp1 subcomplex of Cleavage Factor IA (CF IA), which functions in both 3′ end processing and Pol II termination, and for proper termination of sho… Show more

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Cited by 15 publications
(14 citation statements)
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“…6A) before and after 4-NQO treatment in wild-type and CFIA mutant ( rna15-1 ) strains. The Pol II (Rpb1) occupancy on PMA1 matched previously reported ChIP profiles (Pearson & Moore, 2014), with highest Pol II levels near the 5’-end of the gene and gradual tapering off near the poly(A) site due to transcription termination (Fig. 6B, 6C, top panel ).…”
Section: Resultssupporting
confidence: 85%
See 1 more Smart Citation
“…6A) before and after 4-NQO treatment in wild-type and CFIA mutant ( rna15-1 ) strains. The Pol II (Rpb1) occupancy on PMA1 matched previously reported ChIP profiles (Pearson & Moore, 2014), with highest Pol II levels near the 5’-end of the gene and gradual tapering off near the poly(A) site due to transcription termination (Fig. 6B, 6C, top panel ).…”
Section: Resultssupporting
confidence: 85%
“…Cells were fixed with 1% formaldehyde for 15 minutes and the remaining procedure was performed as described previously using anti-Rpb1 (4H8; SCBT) and anti-Myc (SCBT) antibodies for immunoprecipitation (Pearson & Moore, 2014). …”
Section: Methodsmentioning
confidence: 99%
“…Pcf11, in complex with another CPF-CF subunit, Clp1, has also been shown to bind to the body of the polymerase by interacting with the flap loop domain of the second largest sub unit of Pol II, Rpb2 (REF. 9). It is commonly accepted that both sequences on the nascent RNA and the interaction with the polymerase contribute redundantly to the recruitment of the CPF-CF complex at the 3ʹ end of genes, although the relative contribution of each remains unclear.…”
Section: Poly(a) Site Tss Transcription Unitmentioning
confidence: 93%
“…Extracts were prepared as described by Pearson and Moore ( 69 ) and used to transcribe 32 P-labeled RNA from the DNA templates, which contains a promoter for RNA Pol II and five G-less cassettes of different lengths downstream of the transcription start site, with or without the CYC1 polyA site inserted between the second and third cassettes. RNA produced by transcription was digested with RNase T1, and the RNA fragments were resolved on a 6% polyacrylamide/7 M urea gel and visualized by phosphorimager analysis.…”
Section: Methodsmentioning
confidence: 99%