Cap completion and C-terminal repeat domain kinase recruitment underlie the initiation-elongation transition of RNA polymerase II

Mol Cell Biol. 2013 Oct;33(19):3805-16. doi: 10.1128/MCB.00361-13. Epub 2013 Jul 22.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acid Anhydride Hydrolases / genetics
  • Acid Anhydride Hydrolases / metabolism
  • Chromatin Immunoprecipitation
  • Chromosomal Proteins, Non-Histone / genetics
  • Chromosomal Proteins, Non-Histone / metabolism
  • Cyclin-Dependent Kinases / genetics
  • Cyclin-Dependent Kinases / metabolism
  • Methyltransferases / genetics
  • Methyltransferases / metabolism
  • Models, Genetic
  • Nucleotidyltransferases / genetics
  • Nucleotidyltransferases / metabolism*
  • Protein Binding
  • Protein Kinases / genetics
  • Protein Kinases / metabolism
  • RNA Caps / genetics
  • RNA Caps / metabolism*
  • RNA Polymerase II / metabolism*
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / metabolism
  • Saccharomyces cerevisiae Proteins / genetics
  • Saccharomyces cerevisiae Proteins / metabolism*
  • Transcription Elongation, Genetic*
  • Transcription Initiation Site
  • Transcription Initiation, Genetic*
  • Transcriptional Elongation Factors / genetics
  • Transcriptional Elongation Factors / metabolism

Substances

  • CTDK-I protein complex, S cerevisiae
  • Chromosomal Proteins, Non-Histone
  • RNA Caps
  • Saccharomyces cerevisiae Proteins
  • Transcriptional Elongation Factors
  • SPT5 transcriptional elongation factor
  • Methyltransferases
  • mRNA (guanine(N7))-methyltransferase
  • Protein Kinases
  • Cyclin-Dependent Kinases
  • SGV1 protein, S cerevisiae
  • Nucleotidyltransferases
  • RNA Polymerase II
  • mRNA guanylyltransferase
  • Acid Anhydride Hydrolases
  • RNA triphosphatase