title: A role for CF1A 3' end processing complex in promoter-associated transcription.
abstract: The Cleavage Factor 1A (CF1A) complex, which is required for the termination of transcription in budding yeast, occupies the 3' end of transcriptionally active genes. We recently demonstrated that CF1A subunits also crosslink to the 5' end of genes during transcription. The presence of CF1A complex at the promoter suggested its possible involvement in the initiation/reinitiation of transcription. To check this possibility, we performed transcription run-on assay, RNAP II-density ChIP and strand-specific RT-PCR analysis in a mutant of CF1A subunit Clp1. As expected, RNAP II read through the termination signal in the temperature-sensitive mutant of clp1 at elevated temperature. The transcription readthrough phenotype was accompanied by a decrease in the density of RNAP II in the vicinity of the promoter region. With the exception of TFIIB and TFIIF, the recruitment of the general transcription factors onto the promoter, however, remained unaffected in the clp1 mutant. These results suggest that the CF1A complex affects the recruitment of RNAP II onto the promoter for reinitiation of transcription. Simultaneously, an increase in synthesis of promoter-initiated divergent antisense transcript was observed in the clp1 mutant, thereby implicating CF1A complex in providing directionality to the promoter-bound polymerase. Chromosome Conformation Capture (3C) analysis revealed a physical interaction of the promoter and terminator regions of a gene in the presence of a functional CF1A complex. Gene looping was completely abolished in the clp1 mutant. On the basis of these results, we propose that the CF1A-dependent recruitment of RNAP II onto the promoter for reinitiation and the regulation of directionality of promoter-associated transcription are accomplished through gene looping.
authors: Department of Biological Sciences, Wayne State University, Detroit, Michigan, United States of America.
authors: Nadra
authors: N
authors: Al Husini
authors: Paul
authors: P
authors: Kudla
authors: Athar
authors: A
authors: Ansari
mesh_terms: D020413:3' Untranslated Regions; D005800:Genes, Fungal; D009154:Mutation; D011401:Promoter Regions, Genetic; D012319:RNA Polymerase II / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics*; D035581:Transcription Factor TFIIB / Q000235:genetics*; D019473:Transcription Factors, TFII / Q000235:genetics; D014158:Transcription, Genetic; D039221:mRNA Cleavage and Polyadenylation Factors / Q000235:genetics*
chemical_list: D020413:3' Untranslated Regions; C517046:Clp1 protein, S cerevisiae; C512637:PCF11 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D035581:Transcription Factor TFIIB; D019473:Transcription Factors, TFII; D039221:mRNA Cleavage and Polyadenylation Factors; C512638:RNA14 protein, S cerevisiae; C512635:RNA15 protein, S cerevisiae; D012319:RNA Polymerase II; C056604:transcription factor TFIIF
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