Heptad-Specific Phosphorylation of RNA Polymerase II CTD

Mol Cell. 2016 Jan 21;61(2):305-14. doi: 10.1016/j.molcel.2015.12.003.

Abstract

The carboxy-terminal domain (CTD) of RNA polymerase II (Pol II) consists of heptad repeats with the consensus motif Y1-S2-P3-T4-S5-P6-S7. Dynamic phosphorylation of the CTD coordinates Pol II progression through the transcription cycle. Here, we use genetic and mass spectrometric approaches to directly detect and map phosphosites along the entire CTD. We confirm phosphorylation of CTD residues Y1, S2, T4, S5, and S7 in mammalian and yeast cells. Although specific phosphorylation signatures dominate, adjacent CTD repeats can be differently phosphorylated, leading to a high variation of coexisting phosphosites in mono- and di-heptad CTD repeats. Inhibition of CDK9 kinase specifically reduces S2 phosphorylation levels within the CTD.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Motifs
  • Amino Acid Sequence
  • Animals
  • Cell Line, Tumor
  • Cyclin-Dependent Kinase 9 / antagonists & inhibitors
  • Cyclin-Dependent Kinase 9 / metabolism
  • Humans
  • Mammals
  • Mass Spectrometry
  • Molecular Sequence Data
  • Peptide Library
  • Phosphorylation
  • RNA Polymerase II / chemistry*
  • RNA Polymerase II / metabolism*
  • RNA-Binding Proteins / chemistry
  • RNA-Binding Proteins / metabolism
  • Repetitive Sequences, Amino Acid
  • Reproducibility of Results
  • Saccharomyces cerevisiae / metabolism
  • Saccharomyces cerevisiae Proteins / chemistry
  • Saccharomyces cerevisiae Proteins / metabolism
  • Structure-Activity Relationship

Substances

  • NGR1 protein, S cerevisiae
  • Peptide Library
  • RNA-Binding Proteins
  • Saccharomyces cerevisiae Proteins
  • Cyclin-Dependent Kinase 9
  • RNA Polymerase II