Conformational flexibility of RNA polymerase III during transcriptional elongation

Carlos Fernandez-Tornero, Bettina Boettcher, Umar Jan Rashid, Ulrich Steuerwald, Beate Floerchinger, Damien P. Devos, Doris Lindner, Christoph W. Mueller

Research output: Contribution to journalArticlepeer-review

Abstract

RNA polymerase (Pol) III is responsible for the transcription of genes encoding small RNAs, including tRNA, 5S rRNA and U6 RNA. Here, we report the electron cryomicroscopy structures of yeast Pol III at 9.9 angstrom resolution and its elongation complex at 16.5 angstrom resolution. Particle sub-classification reveals prominent EM densities for the two Pol III-specific subcomplexes, C31/C82/C34 and C37/ C53, that can be interpreted using homology models. While the winged-helix-containing C31/C82/C34 subcomplex initiates transcription from one side of the DNA-binding cleft, the C37/C53 subcomplex accesses the transcription bubble from the opposite side of this cleft. The transcribing Pol III enzyme structure not only shows the complete incoming DNA duplex, but also reveals the exit path of newly synthesized RNA. During transcriptional elongation, the Pol III-specific subcomplexes tightly enclose the incoming DNA duplex, which likely increases processivity and provides structural insights into the conformational switch between Pol III-mediated initiation and elongation.

Original languageEnglish
Pages (from-to)3762-3772
Number of pages11
JournalEMBO Journal
Volume29
Issue number22
DOIs
Publication statusPublished - 17 Nov 2010

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