Architecture and RNA binding of the human negative elongation factor
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...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
eLife Sciences Publications Ltd
2016-06-01
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| Series: | eLife |
| Subjects: | |
| Online Access: | https://elifesciences.org/articles/14981 |
| _version_ | 1811180438706716672 |
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| author | Seychelle M Vos David Pöllmann Livia Caizzi Katharina B Hofmann Pascaline Rombaut Tomasz Zimniak Franz Herzog Patrick Cramer |
| author_facet | Seychelle M Vos David Pöllmann Livia Caizzi Katharina B Hofmann Pascaline Rombaut Tomasz Zimniak Franz Herzog Patrick Cramer |
| author_sort | Seychelle M Vos |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-04-11T09:03:10Z |
| format | Article |
| id | doaj.art-caf7bfc79a114ab4a6e1952f7a1bedf4 |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-11T09:03:10Z |
| publishDate | 2016-06-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-caf7bfc79a114ab4a6e1952f7a1bedf42022-12-22T04:32:43ZengeLife Sciences Publications LtdeLife2050-084X2016-06-01510.7554/eLife.14981Architecture and RNA binding of the human negative elongation factorSeychelle M Vos0David Pöllmann1Livia Caizzi2Katharina B Hofmann3Pascaline Rombaut4Tomasz Zimniak5Franz Herzog6Patrick Cramer7https://orcid.org/0000-0001-5454-7755Department of Molecular Biology, Max Planck Institute for Biophysical Chemistry, Göttingen, GermanyDepartment of Molecular Biology, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany; Gene Center Munich, Ludwig-Maximilians-Universität München, Munich, Germany; Department of Biochemistry, Ludwig-Maximilians-Universität München, Munich, GermanyDepartment of Molecular Biology, Max Planck Institute for Biophysical Chemistry, Göttingen, GermanyDepartment of Molecular Biology, Max Planck Institute for Biophysical Chemistry, Göttingen, GermanyGene Center Munich, Ludwig-Maximilians-Universität München, Munich, Germany; Department of Biochemistry, Ludwig-Maximilians-Universität München, Munich, GermanyGene Center Munich, Ludwig-Maximilians-Universität München, Munich, Germany; Department of Biochemistry, Ludwig-Maximilians-Universität München, Munich, GermanyGene Center Munich, Ludwig-Maximilians-Universität München, Munich, Germany; Department of Biochemistry, Ludwig-Maximilians-Universität München, Munich, GermanyDepartment of Molecular Biology, Max Planck Institute for Biophysical Chemistry, Göttingen, GermanyTranscription 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.https://elifesciences.org/articles/14981gene transcriptionRNA polymerase IIgene regulationpromoter-proximal pausing |
| spellingShingle | Seychelle M Vos David Pöllmann Livia Caizzi Katharina B Hofmann Pascaline Rombaut Tomasz Zimniak Franz Herzog Patrick Cramer Architecture and RNA binding of the human negative elongation factor eLife gene transcription RNA polymerase II gene regulation promoter-proximal pausing |
| title | Architecture and RNA binding of the human negative elongation factor |
| title_full | Architecture and RNA binding of the human negative elongation factor |
| title_fullStr | Architecture and RNA binding of the human negative elongation factor |
| title_full_unstemmed | Architecture and RNA binding of the human negative elongation factor |
| title_short | Architecture and RNA binding of the human negative elongation factor |
| title_sort | architecture and rna binding of the human negative elongation factor |
| topic | gene transcription RNA polymerase II gene regulation promoter-proximal pausing |
| url | https://elifesciences.org/articles/14981 |
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