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Title: Structural visualization of RNA polymerase III transcription machineries

Abstract

RNA polymerase III (Pol III) transcription initiation requires the action of the transcription factor IIIB (TFIIIB) and is highly regulated. Here, we determine the structures of Pol III pre-initiation complexes (PICs) using single particle cryo-electron microscopy (cryo-EM). We observe stable Pol III–TFIIIB complexes using nucleic acid scaffolds mimicking various functional states, in which TFIIIB tightly encircles the upstream promoter DNA. There is an intricate interaction between TFIIIB and Pol III, which stabilizes the winged-helix domains of the C34 subunit of Pol III over the active site cleft. The architecture of Pol III PIC more resembles that of the Pol II PIC than the Pol I PIC. In addition, we also obtain a 3D reconstruction of Pol III in complex with TFIIIB using the elongation complex (EC) scaffold, shedding light on the mechanism of facilitated recycling of Pol III prior to transcription re-initiation.

Authors:
ORCiD logo; ; ; ORCiD logo; ORCiD logo
Publication Date:
Research Org.:
Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States). National Energy Research Scientific Computing Center (NERSC); Univ. of California, Oakland, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC)
OSTI Identifier:
1459999
Alternate Identifier(s):
OSTI ID: 1543739
Grant/Contract Number:  
AC02-05CH11231
Resource Type:
Journal Article: Published Article
Journal Name:
Cell Discovery
Additional Journal Information:
Journal Name: Cell Discovery Journal Volume: 4 Journal Issue: 1; Journal ID: ISSN 2056-5968
Publisher:
Nature Publishing Group
Country of Publication:
Country unknown/Code not available
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; Cell Biology

Citation Formats

Han, Yan, Yan, Chunli, Fishbain, Susan, Ivanov, Ivaylo, and He, Yuan. Structural visualization of RNA polymerase III transcription machineries. Country unknown/Code not available: N. p., 2018. Web. doi:10.1038/s41421-018-0044-z.
Han, Yan, Yan, Chunli, Fishbain, Susan, Ivanov, Ivaylo, & He, Yuan. Structural visualization of RNA polymerase III transcription machineries. Country unknown/Code not available. doi:10.1038/s41421-018-0044-z.
Han, Yan, Yan, Chunli, Fishbain, Susan, Ivanov, Ivaylo, and He, Yuan. Tue . "Structural visualization of RNA polymerase III transcription machineries". Country unknown/Code not available. doi:10.1038/s41421-018-0044-z.
@article{osti_1459999,
title = {Structural visualization of RNA polymerase III transcription machineries},
author = {Han, Yan and Yan, Chunli and Fishbain, Susan and Ivanov, Ivaylo and He, Yuan},
abstractNote = {RNA polymerase III (Pol III) transcription initiation requires the action of the transcription factor IIIB (TFIIIB) and is highly regulated. Here, we determine the structures of Pol III pre-initiation complexes (PICs) using single particle cryo-electron microscopy (cryo-EM). We observe stable Pol III–TFIIIB complexes using nucleic acid scaffolds mimicking various functional states, in which TFIIIB tightly encircles the upstream promoter DNA. There is an intricate interaction between TFIIIB and Pol III, which stabilizes the winged-helix domains of the C34 subunit of Pol III over the active site cleft. The architecture of Pol III PIC more resembles that of the Pol II PIC than the Pol I PIC. In addition, we also obtain a 3D reconstruction of Pol III in complex with TFIIIB using the elongation complex (EC) scaffold, shedding light on the mechanism of facilitated recycling of Pol III prior to transcription re-initiation.},
doi = {10.1038/s41421-018-0044-z},
journal = {Cell Discovery},
issn = {2056-5968},
number = 1,
volume = 4,
place = {Country unknown/Code not available},
year = {2018},
month = {7}
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record at 10.1038/s41421-018-0044-z

Citation Metrics:
Cited by: 2 works
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Figures / Tables:

Fig. 1 Fig. 1: Cryo-EM structure of the Pol III ITC. a Nucleic acid scaffold used. The non-template (top) and template (bottom) strands are depicted in cyan. Mismatched sequence is shown as separate strands. RNA is shown in red. TATA box is depicted in yellow. TFIIIB binding region and transcription bubble positionmore » are also labeled. b Cryo-EM reconstruction of the Pol III ITC. Pol III is colored gray, and the nucleic acid template is colored cyan. The TFIIIB subunits are depicted in blue (Brf1), red (TBP), and orange (Bdp1). Two views, front (left) and bottom (right), are shown. c Two views of the Pol III ITC model. Components are colored the same as in b« less

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    Works referencing / citing this record:

    The multi-faceted regulation of nuclear tRNA gene transcription: Transfer RNA gene expression
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    • Hummel, Guillaume; Warren, Jessica; Drouard, Laurence
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