ORCID: https://orcid.org/0000-0001-8270-1449 and Cramer, Patrick
(2016):
Architecture and RNA binding of the human negative elongation factor.
In: eLife, Vol. 5, e14981
[PDF, 11MB]


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.
Item Type: | Journal article |
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EU Funded Grant Agreement Number: | 638218 |
EU Projects: | Horizon 2020 > ERC Grants > ERC Starting Grant > ERC Grant 638218: MolStruKT - Molecular structure and cell cycle regulated assembly of the kinetochore |
Faculties: | Chemistry and Pharmacy > Department of Biochemistry Chemistry and Pharmacy > GeneCenter |
Subjects: | 500 Science > 540 Chemistry |
URN: | urn:nbn:de:bvb:19-epub-76124-5 |
Language: | English |
Item ID: | 76124 |
Date Deposited: | 01. Jun 2021 07:54 |
Last Modified: | 01. Jun 2021 10:45 |