Exchange of RNA polymerase II initiation and elongation factors during gene expression in vivo.
We have systematically explored the in vivo occupancy of promoters and open reading frames by components of the RNA polymerase II transcription initiation and elongation apparatuses in yeast. RNA polymerase II, Mediator, and the general transcription factors (GTFs) were recruited to all promoters tested upon gene activation. RNA polymerase II, TFIIS, Spt5, and, unexpectedly, the Paf1/Cdc73 complex, were found associated with open reading frames. The presence of the Paf1/Cdc73 complex on ORFs in vivo suggests a novel function for this complex in elongation. Elongator was not detected under any conditions tested, and further analysis revealed that the majority of elongator is cytoplasmic. These results suggest a revised model for transcription initiation and elongation apparatuses in living cells.
Pokholok
Dmitry K
DK
Whitehead Institute for Biomedical Research, 9 Cambridge Center, Cambridge, MA 02142, USA.
Hannett
Nancy M
NM
Young
Richard A
RA
D002843:Chromatin / Q000737:chemistry; D002868:Chromosomal Proteins, Non-Histone; D004271:DNA, Fungal / Q000235:genetics / Q000378:metabolism; D005686:Galactokinase / Q000235:genetics; D015966:Gene Expression Regulation, Fungal; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics; D018841:HSP90 Heat-Shock Proteins; D006360:Heat-Shock Proteins / Q000235:genetics; D006358:Hot Temperature; D046911:Macromolecular Substances; D008957:Models, Genetic; D009687:Nuclear Proteins / Q000378:metabolism*; D016366:Open Reading Frames; D011233:Precipitin Tests; D011401:Promoter Regions, Genetic; D012319:RNA Polymerase II / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D014157:Transcription Factors / Q000378:metabolism*; D035165:Transcription Factors, General; D014158:Transcription, Genetic; D015533:Transcriptional Activation; D035602:Transcriptional Elongation Factors
C104439:CDC73 protein, S cerevisiae; D002843:Chromatin; D002868:Chromosomal Proteins, Non-Histone; D004271:DNA, Fungal; D018840:HSP70 Heat-Shock Proteins; C074510:HSP82 protein, S cerevisiae; D018841:HSP90 Heat-Shock Proteins; D006360:Heat-Shock Proteins; D046911:Macromolecular Substances; D009687:Nuclear Proteins; C097698:PAF1 protein, S cerevisiae; D011993:Recombinant Fusion Proteins; C456269:SSA4 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D035165:Transcription Factors, General; D035602:Transcriptional Elongation Factors; C018648:transcription factor S-II; C069940:SPT5 transcriptional elongation factor; D005686:Galactokinase; D012319:RNA Polymerase II