Architecture and RNA binding of the human negative elongation factor
| Autor: | Livia Caizzi, Franz Herzog, Tomasz Zimniak, David Pöllmann, Patrick Cramer, Pascaline Rombaut, Seychelle M. Vos, Katharina B Hofmann |
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| Jazyk: | angličtina |
| Předmět: |
0301 basic medicine
Models Molecular RNA polymerase II biochemistry biophysics gene regulation gene transcription human promoter-proximal pausing structural biology Transcription Genetic QH301-705.5 Protein Conformation Science RNA-dependent RNA polymerase macromolecular substances Biology Crystallography X-Ray Biochemistry General Biochemistry Genetics and Molecular Biology 03 medical and health sciences Transcription (biology) RNA polymerase I Humans Biology (General) Negative elongation factor General Immunology and Microbiology General Neuroscience RNA General Medicine Biophysics and Structural Biology Molecular biology Cell biology RNA silencing 030104 developmental biology Gene Expression Regulation Medicine Transcription factor II D Small nuclear RNA Research Article Human Protein Binding Transcription Factors |
| Zdroj: | eLife eLife, Vol 5 (2016) |
| ISSN: | 2050-084X |
| DOI: | 10.7554/elife.14981 |
| Popis: | 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. DOI: http://dx.doi.org/10.7554/eLife.14981.001 |
| Databáze: | OpenAIRE |
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