title: Two RNA polymerase I subunits control the binding and release of Rrn3 during transcription.
abstract: Rpa34 and Rpa49 are nonessential subunits of RNA polymerase I, conserved in species from Saccharomyces cerevisiae and Schizosaccharomyces pombe to humans. Rpa34 bound an N-terminal region of Rpa49 in a two-hybrid assay and was lost from RNA polymerase in an rpa49 mutant lacking this Rpa34-binding domain, whereas rpa34Delta weakened the binding of Rpa49 to RNA polymerase. rpa34Delta mutants were caffeine sensitive, and the rpa34Delta mutation was lethal in a top1Delta mutant and in rpa14Delta, rpa135(L656P), and rpa135(D395N) RNA polymerase mutants. These defects were shared by rpa49Delta mutants, were suppressed by the overexpression of Rpa49, and thus, were presumably mediated by Rpa49 itself. rpa49 mutants lacking the Rpa34-binding domain behaved essentially like rpa34Delta mutants, but strains carrying rpa49Delta and rpa49-338::HIS3 (encoding a form of Rpa49 lacking the conserved C terminus) had reduced polymerase occupancy at 30 degrees C, failed to grow at 25 degrees C, and were sensitive to 6-azauracil and mycophenolate. Mycophenolate almost fully dissociated the mutant polymerase from its ribosomal DNA (rDNA) template. The rpa49Delta and rpa49-338::HIS3 mutations had a dual effect on the transcription initiation factor Rrn3 (TIF-IA). They partially impaired its recruitment to the rDNA promoter, an effect that was bypassed by an N-terminal deletion of the Rpa43 subunit encoded by rpa43-35,326, and they strongly reduced the release of the Rrn3 initiation factor during elongation. These data suggest a dual role of the Rpa49-Rpa34 dimer during the recruitment of Rrn3 and its subsequent dissociation from the elongating polymerase.
authors: CEA, IbiTec-S, Service de Biologie Intégrative & Génétique Moléculaire, Gif sur Yvette Cedex F-91191, France.
authors: Frédéric
authors: F
authors: Beckouet
authors: Sylvie
authors: S
authors: Labarre-Mariotte
authors: Benjamin
authors: B
authors: Albert
authors: Yukiko
authors: Y
authors: Imazawa
authors: Michel
authors: M
authors: Werner
authors: Olivier
authors: O
authors: Gadal
authors: Yasuhisa
authors: Y
authors: Nogi
authors: Pierre
authors: P
authors: Thuriaux
mesh_terms: D000595:Amino Acid Sequence; D047369:Chromatin Immunoprecipitation; D019281:Dimerization; D004791:Enzyme Inhibitors / Q000494:pharmacology; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation; D009173:Mycophenolic Acid / Q000494:pharmacology; D010957:Plasmids; D035463:Pol1 Transcription Initiation Complex Proteins / Q000378:metabolism*; D011485:Protein Binding; D021122:Protein Subunits / Q000235:genetics / Q000378:metabolism*; D012318:RNA Polymerase I / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism*; D012568:Schizosaccharomyces / Q000201:enzymology / Q000235:genetics / Q000378:metabolism; D029702:Schizosaccharomyces pombe Proteins / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D014157:Transcription Factors / Q000378:metabolism*; D014158:Transcription, Genetic; D020798:Two-Hybrid System Techniques
chemical_list: D004791:Enzyme Inhibitors; D035463:Pol1 Transcription Initiation Complex Proteins; D021122:Protein Subunits; C101161:RRN3 protein, S cerevisiae; C527303:Rrn3 protein, S pombe; D029701:Saccharomyces cerevisiae Proteins; D029702:Schizosaccharomyces pombe Proteins; D014157:Transcription Factors; D012318:RNA Polymerase I; D009173:Mycophenolic Acid