Architecture and RNA binding of the human negative elongation factor

Vos, Seychelle M., Pöllmann, David, Caizzi, Livia, Hofmann, Katharina B., Rombaut, Pascaline, Zimniak, Tomasz, Herzog, Franz and Cramer, Patrick (2016): Architecture and RNA binding of the human negative elongation factor. In: eLife, Vol. 5, e14981

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
• 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
• Language: English