Methods review: Mass spectrometry analysis of RNAPII complexes

Katlyn Hughes Burriss, Amber L. Mosley

Research output: Contribution to journalReview article

2 Scopus citations

Abstract

RNA Polymerase II (RNAPII) is responsible for transcribing multiple RNA species throughout eukaryotes. A variety of protein-protein interactions occur throughout the transcription cycle for coordinated regulation of transcription initiation, elongation, and/or termination. Taking a proteomics approach to study RNAPII transcription thereby offers a comprehensive view of both RNAPII biology and the variety of proteins that regulate the process itself. This review will focus on how mass spectrometry (MS) methods have expanded understanding of RNAPII and its transcription-regulatory interaction partners. The application of affinity purification mass spectrometry has led to the discovery of a number of novel groups of proteins that regulate an array of RNAPII biology ranging from nuclear import to regulation of phosphorylation state. Additionally, a number of methods have been developed using mass spectrometry to measure protein subunit stoichiometry within and across protein complexes and to perform various types of architectural analysis using structural proteomics approaches. The key methods that we will focus on related to RNAPII mass spectrometry analyses include: affinity purification mass spectrometry, protein post-translational modification analysis, crosslinking mass spectrometry, and native mass spectrometry.

Original languageEnglish (US)
Pages (from-to)105-114
Number of pages10
JournalMethods
Volume159-160
DOIs
StatePublished - Apr 15 2019

Keywords

  • Affinity purification
  • C-terminal domain
  • CTD
  • Crosslinking mass spectrometry
  • Kinase
  • Mass spectrometry
  • Nano-electrospray ionization
  • Non-covalent interaction networks
  • Phosphorylation
  • Proteomics
  • RNA Polymerase II
  • Structural proteomics
  • Transcription

ASJC Scopus subject areas

  • Molecular Biology
  • Biochemistry, Genetics and Molecular Biology(all)

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