Architecture of the RNA polymerase II-Paf1C-TFIIS transcription elongation complex

Youwei Xu; Carrie Bernecky; Chung-Tien Lee; Kerstin C Maier; Björn Schwalb; Dimitry Tegunov; Jürgen M Plitzko; Henning Urlaub; Patrick Cramer

doi:10.1038/ncomms15741

Architecture of the RNA polymerase II-Paf1C-TFIIS transcription elongation complex

Nat Commun. 2017 Jun 6:8:15741. doi: 10.1038/ncomms15741.

Authors

Affiliations

¹ Department of Molecular Biology, Max-Planck-Institute for Biophysical Chemistry, Max Planck Society, Am Fassberg 11, Göttingen 37077, Germany.
² Bioanalytical Mass Spectrometry, Max-Planck-Institute for Biophysical Chemistry, Am Fassberg 11, Göttingen 37077, Germany.
³ Bioanalytics Group, Institute for Clinical Chemistry, University Medical Center, Göttingen, Robert-Koch-Strasse 40, Göttingen 37075, Germany.
⁴ Department of Molecular Structural Biology, Max-Planck-Institute for Biochemistry, Am Klopferspitz 18, Martinsried 82152, Germany.

Abstract

The conserved polymerase-associated factor 1 complex (Paf1C) plays multiple roles in chromatin transcription and genomic regulation. Paf1C comprises the five subunits Paf1, Leo1, Ctr9, Cdc73 and Rtf1, and binds to the RNA polymerase II (Pol II) transcription elongation complex (EC). Here we report the reconstitution of Paf1C from Saccharomyces cerevisiae, and a structural analysis of Paf1C bound to a Pol II EC containing the elongation factor TFIIS. Cryo-electron microscopy and crosslinking data reveal that Paf1C is highly mobile and extends over the outer Pol II surface from the Rpb2 to the Rpb3 subunit. The Paf1-Leo1 heterodimer and Cdc73 form opposite ends of Paf1C, whereas Ctr9 bridges between them. Consistent with the structural observations, the initiation factor TFIIF impairs Paf1C binding to Pol II, whereas the elongation factor TFIIS enhances it. We further show that Paf1C is globally required for normal mRNA transcription in yeast. These results provide a three-dimensional framework for further analysis of Paf1C function in transcription through chromatin.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Binding, Competitive
Cell Cycle Proteins / chemistry
Cell Cycle Proteins / genetics
Cell Cycle Proteins / metabolism
Cross-Linking Reagents / chemistry
Cryoelectron Microscopy
Multiprotein Complexes / chemistry*
Multiprotein Complexes / metabolism
Nuclear Proteins / chemistry*
Nuclear Proteins / genetics
Nuclear Proteins / metabolism
Protein Conformation
RNA Polymerase II / chemistry*
RNA Polymerase II / genetics
RNA Polymerase II / metabolism*
RNA-Binding Proteins / chemistry
RNA-Binding Proteins / metabolism
Recombinant Proteins / chemistry
Recombinant Proteins / genetics
Recombinant Proteins / metabolism
Saccharomyces cerevisiae Proteins / chemistry*
Saccharomyces cerevisiae Proteins / genetics
Saccharomyces cerevisiae Proteins / metabolism
Transcription, Genetic
Transcriptional Elongation Factors / chemistry*
Transcriptional Elongation Factors / genetics
Transcriptional Elongation Factors / metabolism

Substances

CTR9 protein, S cerevisiae
Cell Cycle Proteins
Cross-Linking Reagents
Leo1 protein, S cerevisiae
Multiprotein Complexes
Nuclear Proteins
PAF1 protein, S cerevisiae
RNA-Binding Proteins
Recombinant Proteins
Saccharomyces cerevisiae Proteins
Transcriptional Elongation Factors
transcription factor S-II
RNA Polymerase II
RPB2 protein, S cerevisiae
RPB3 protein, S cerevisiae