In:
eLife, eLife Sciences Publications, Ltd, Vol. 5 ( 2016-06-10)
Abstract:
Transcription regulation in metazoans often involves promoter-proximal pausing of RNA polymerase (Pol) II, which requires the 4-subunit negative elongation factor (NELF). Here we discern the functional architecture of human NELF through X-ray crystallography, protein crosslinking, biochemical assays, and RNA crosslinking in cells. We identify a NELF core subcomplex formed by conserved regions in subunits NELF-A and NELF-C, and resolve its crystal structure. The NELF-AC subcomplex binds single-stranded nucleic acids in vitro, and NELF-C associates with RNA in vivo. A positively charged face of NELF-AC is involved in RNA binding, whereas the opposite face of the NELF-AC subcomplex binds NELF-B. NELF-B is predicted to form a HEAT repeat fold, also binds RNA in vivo, and anchors the subunit NELF-E, which is confirmed to bind RNA in vivo. These results reveal the three-dimensional architecture and three RNA-binding faces of NELF.
Type of Medium:
Online Resource
ISSN:
2050-084X
DOI:
10.7554/eLife.14981.001
DOI:
10.7554/eLife.14981.002
DOI:
10.7554/eLife.14981.003
DOI:
10.7554/eLife.14981.004
DOI:
10.7554/eLife.14981.005
DOI:
10.7554/eLife.14981.006
DOI:
10.7554/eLife.14981.007
DOI:
10.7554/eLife.14981.008
DOI:
10.7554/eLife.14981.009
DOI:
10.7554/eLife.14981.010
DOI:
10.7554/eLife.14981.011
DOI:
10.7554/eLife.14981.012
DOI:
10.7554/eLife.14981.013
DOI:
10.7554/eLife.14981.014
DOI:
10.7554/eLife.14981.015
DOI:
10.7554/eLife.14981.016
DOI:
10.7554/eLife.14981.017
DOI:
10.7554/eLife.14981.018
DOI:
10.7554/eLife.14981.019
DOI:
10.7554/eLife.14981.020
DOI:
10.7554/eLife.14981.021
DOI:
10.7554/eLife.14981.022
DOI:
10.7554/eLife.14981.023
DOI:
10.7554/eLife.14981.024
DOI:
10.7554/eLife.14981.026
DOI:
10.7554/eLife.14981.027
Language:
English
Publisher:
eLife Sciences Publications, Ltd
Publication Date:
2016
detail.hit.zdb_id:
2687154-3
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