title: Cap completion and C-terminal repeat domain kinase recruitment underlie the initiation-elongation transition of RNA polymerase II.
abstract: After transcription initiation, RNA polymerase (Pol) II escapes from the promoter and recruits elongation factors. The molecular basis for the initiation-elongation factor exchange during this transition remains poorly understood. Here, we used chromatin immunoprecipitation (ChIP) to elucidate the initiation-elongation transition of Pol II in the budding yeast Saccharomyces cerevisiae. We show that the early Pol II elongation factor Spt5 contributes to stable recruitment of the mRNA capping enzymes Cet1, Ceg1, and Abd1. Genome-wide occupancy for Cet1 and Ceg1 is restricted to the transcription start site (TSS), whereas occupancy for Abd1 peaks at ~110 nucleotides downstream, and occupancy for the cap-binding complex (CBC) rises subsequently. Abd1 and CBC are important for recruitment of the kinases Ctk1 and Bur1, which promote elongation and capping enzyme release. These results suggest that cap completion stimulates productive Pol II elongation.
authors.affiliation: Gene Center and Department of Biochemistry, Center for Integrated Protein Science Munich, Ludwig Maximilians Universität München, Munich, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Lidschreiber
authors.forename: Kristin
authors.initials: K
authors.lastname: Leike
authors.forename: Patrick
authors.initials: P
authors.lastname: Cramer
mesh_terms: D017766:Acid Anhydride Hydrolases / Q000235:genetics / Q000378:metabolism; D047369:Chromatin Immunoprecipitation; D002868:Chromosomal Proteins, Non-Histone / Q000235:genetics / Q000378:metabolism; D018844:Cyclin-Dependent Kinases / Q000235:genetics / Q000378:metabolism; D008780:Methyltransferases / Q000235:genetics / Q000378:metabolism; D008957:Models, Genetic; D009713:Nucleotidyltransferases / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D011494:Protein Kinases / Q000235:genetics / Q000378:metabolism; D012315:RNA Caps / Q000235:genetics / Q000378:metabolism*; D012319:RNA Polymerase II / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D061805:Transcription Elongation, Genetic; D024363:Transcription Initiation Site; D061785:Transcription Initiation, Genetic; D035602:Transcriptional Elongation Factors / Q000235:genetics / Q000378:metabolism; D000098556:mRNA Guanylyltransferases
chemical_list: C118305:CTDK-I protein complex, S cerevisiae; D002868:Chromosomal Proteins, Non-Histone; D012315:RNA Caps; D029701:Saccharomyces cerevisiae Proteins; D035602:Transcriptional Elongation Factors; C069940:SPT5 transcriptional elongation factor; D008780:Methyltransferases; C055097:mRNA (guanine(N7))-methyltransferase; D011494:Protein Kinases; D018844:Cyclin-Dependent Kinases; C414163:SGV1 protein, S cerevisiae; D009713:Nucleotidyltransferases; D012319:RNA Polymerase II; D000098556:mRNA Guanylyltransferases; D017766:Acid Anhydride Hydrolases; C023090:RNA triphosphatase