title: Evidence for a complex of transcription factor IIB with poly(A) polymerase and cleavage factor 1 subunits required for gene looping.
abstract: Gene looping, defined as the interaction of the promoter and the terminator regions of a gene during transcription, requires transcription factor IIB (TFIIB). We have earlier demonstrated association of TFIIB with the distal ends of a gene in an activator-dependent manner (El Kaderi, B., Medler, S., Raghunayakula, S., and Ansari, A. (2009) J. Biol. Chem. 284, 25015-25025). The presence of TFIIB at the 3' end of a gene required its interaction with cleavage factor 1 (CF1) 3' end processing complex subunit Rna15. Here, employing affinity chromatography and glycerol gradient centrifugation, we show that TFIIB associates with poly(A) polymerase and the entire CF1 complex in yeast cells. The factors required for general transcription such as TATA-binding protein, RNA polymerase II, and TFIIH are not a component of the TFIIB complex. This holo-TFIIB complex was resistant to MNase digestion. The complex was observed only in the looping-competent strains, but not in the looping-defective sua7-1 strain. The requirement of Rna15 in gene looping has been demonstrated earlier. Here we provide evidence that poly(A) polymerase (Pap1) as well as CF1 subunits Rna14 and Pcf11 are also required for loop formation of MET16 and INO1 genes. Accordingly, cross-linking of TFIIB to the 3' end of genes was abolished in the mutants of Pap1, Rna14, and Pcf11. We further show that in sua7-1 cells, where holo-TFIIB complex is not formed, the kinetics of activated transcription is altered. These results suggest that a complex of TFIIB, CF1 subunits, and Pap1 exists in yeast cells. Furthermore, TFIIB interaction with the CF1 complex and Pap1 is crucial for gene looping and transcriptional regulation.
authors.affiliation: Department of Biological Science, Wayne State University, Detroit, Michigan 48202, USA.
authors.forename: Scott
authors.initials: S
authors.lastname: Medler
authors.forename: Nadra
authors.initials: N
authors.lastname: Al Husini
authors.forename: Sarita
authors.initials: S
authors.lastname: Raghunayakula
authors.forename: Banupriya
authors.initials: B
authors.lastname: Mukundan
authors.forename: Ashley
authors.initials: A
authors.lastname: Aldea
authors.forename: Athar
authors.initials: A
authors.lastname: Ansari
mesh_terms: D020413:3' Untranslated Regions / Q000502:physiology; D002846:Chromatography, Affinity; D004271:DNA, Fungal / Q000235:genetics / Q000378:metabolism; D005800:Genes, Fungal / Q000502:physiology; D046912:Multiprotein Complexes / Q000235:genetics / Q000378:metabolism*; D009154:Mutation; D007296:Myo-Inositol-1-Phosphate Synthase / Q000235:genetics / Q000378:metabolism; D010088:Oxidoreductases / Q000235:genetics / Q000378:metabolism; D000073718:Pancreatitis-Associated Proteins; D011062:Polynucleotide Adenylyltransferase / Q000235:genetics / Q000378:metabolism*; D012319:RNA Polymerase II / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D035581:Transcription Factor TFIIB / Q000235:genetics / Q000378:metabolism*; D039221:mRNA Cleavage and Polyadenylation Factors / Q000235:genetics / Q000378:metabolism*
chemical_list: D020413:3' Untranslated Regions; D004271:DNA, Fungal; D046912:Multiprotein Complexes; C512637:PCF11 protein, S cerevisiae; D000073718:Pancreatitis-Associated Proteins; C000612141:REG3A protein, human; C073680:SUA7 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D035581:Transcription Factor TFIIB; D039221:mRNA Cleavage and Polyadenylation Factors; C512638:RNA14 protein, S cerevisiae; C512635:RNA15 protein, S cerevisiae; C508234:MET16 protein, S cerevisiae; D010088:Oxidoreductases; D012319:RNA Polymerase II; C429559:PAP1 protein, S cerevisiae; D011062:Polynucleotide Adenylyltransferase; C519641:INO1 protein, S cerevisiae; D007296:Myo-Inositol-1-Phosphate Synthase