Schulz, Sarah and Gietl, Andreas and Smollett, Katherine and Tinnefeld, Philip and Werner, Finn and Grohmann, Dina (2016) TFE and Spt4/5 open and close the RNA polymerase clamp during the transcription cycle. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 113 (13). E1816-E1825. ISSN 0027-8424,
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Transcription is an intrinsically dynamic process and requires the coordinated interplay of RNA polymerases (RNAPs) with nucleic acids and transcription factors. Classical structural biology techniques have revealed detailed snapshots of a subset of conformational states of the RNAP as they exist in crystals. A detailed view of the conformational space sampled by the RNAP and the molecular mechanisms of the basal transcription factors E (TFE) and Spt4/5 through conformational constraints has remained elusive. We monitored the conformational changes of the flexible clamp of the RNAP by combining a fluorescently labeled recombinant 12-subunit RNAP system with single-molecule FRET measurements. We measured and compared the distances across the DNA binding channel of the archaeal RNAP. Our results show that the transition of the closed to the open initiation complex, which occurs concomitant with DNA melting, is coordinated with an opening of the RNAP clamp that is stimulated by TFE. We show that the clamp in elongation complexes is modulated by the nontemplate strand and by the processivity factor Spt4/5, both of which stimulate transcription processivity. Taken together, our results reveal an intricate network of interactions within transcription complexes between RNAP, transcription factors, and nucleic acids that allosterically modulate the RNAP during the transcription cycle.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | MOLECULE FLUORESCENCE SPECTROSCOPY; II ELONGATION COMPLEX; RAY CRYSTAL-STRUCTURE; OPEN PROMOTER COMPLEX; STRUCTURAL BASIS; PREINITIATION COMPLEX; ARCHAEAL TRANSCRIPTION; ANGSTROM RESOLUTION; ESCHERICHIA-COLI; SWITCH REGION; RNA polymerase; single-molecule FRET; transcription; archaea; clamp |
| Subjects: | 500 Science > 570 Life sciences |
| Divisions: | Biology, Preclinical Medicine > Institut für Biochemie, Genetik und Mikrobiologie > Lehrstuhl für Mikrobiologie > Prof. Dr. Dina Grohmann |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 03 Apr 2019 10:22 |
| Last Modified: | 03 Apr 2019 10:22 |
| URI: | https://pred.uni-regensburg.de/id/eprint/3221 |
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