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 and Patrick Cramer ()
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Youwei Xu: Max-Planck-Institute for Biophysical Chemistry, Max Planck Society
Carrie Bernecky: Max-Planck-Institute for Biophysical Chemistry, Max Planck Society
Chung-Tien Lee: Bioanalytical Mass Spectrometry, Max-Planck-Institute for Biophysical Chemistry
Kerstin C. Maier: Max-Planck-Institute for Biophysical Chemistry, Max Planck Society
Björn Schwalb: Max-Planck-Institute for Biophysical Chemistry, Max Planck Society
Dimitry Tegunov: Max-Planck-Institute for Biophysical Chemistry, Max Planck Society
Jürgen M. Plitzko: Max-Planck-Institute for Biochemistry
Henning Urlaub: Bioanalytical Mass Spectrometry, Max-Planck-Institute for Biophysical Chemistry
Patrick Cramer: Max-Planck-Institute for Biophysical Chemistry, Max Planck Society

Nature Communications, 2017, vol. 8, issue 1, 1-13

Abstract: 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.

Date: 2017
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DOI: 10.1038/ncomms15741

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