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Structure paper

TitleStructure of activated transcription complex Pol II-DSIF-PAF-SPT6.
Journal, issue, pagesNature, Vol. 560, Issue 7720, Page 607-612, Year 2018
Publish dateAug 22, 2018
AuthorsSeychelle M Vos / Lucas Farnung / Marc Boehning / Christoph Wigge / Andreas Linden / Henning Urlaub / Patrick Cramer /
PubMed AbstractGene regulation involves activation of RNA polymerase II (Pol II) that is paused and bound by the protein complexes DRB sensitivity-inducing factor (DSIF) and negative elongation factor (NELF). Here ...Gene regulation involves activation of RNA polymerase II (Pol II) that is paused and bound by the protein complexes DRB sensitivity-inducing factor (DSIF) and negative elongation factor (NELF). Here we show that formation of an activated Pol II elongation complex in vitro requires the kinase function of the positive transcription elongation factor b (P-TEFb) and the elongation factors PAF1 complex (PAF) and SPT6. The cryo-EM structure of an activated elongation complex of Sus scrofa Pol II and Homo sapiens DSIF, PAF and SPT6 was determined at 3.1 Å resolution and compared to the structure of the paused elongation complex formed by Pol II, DSIF and NELF. PAF displaces NELF from the Pol II funnel for pause release. P-TEFb phosphorylates the Pol II linker to the C-terminal domain. SPT6 binds to the phosphorylated C-terminal-domain linker and opens the RNA clamp formed by DSIF. These results provide the molecular basis for Pol II pause release and elongation activation.
External linksNature / PubMed:30135578
MethodsEM (single particle) / X-ray diffraction
Resolution1.802 - 3.49 Å
Structure data

EMDB-0030:
Structure of activated transcription complex Pol II-DSIF-PAF-SPT6 (Map B, NGN/SPT4 selected particles)
Method: EM (single particle) / Resolution: 3.34 Å

EMDB-0031: Structure of activated transcription complex Pol II-DSIF-PAF-SPT6 (Map A)
PDB-6gmh: Structure of activated transcription complex Pol II-DSIF-PAF-SPT6
Method: EM (single particle) / Resolution: 3.1 Å

EMDB-0032:
Structure of activated transcription complex Pol II-DSIF-PAF-SPT6, KOW1 selected particles (Map C)
Method: EM (single particle) / Resolution: 3.49 Å

EMDB-0033:
Structure of activated transcription complex Pol II-DSIF-PAF-SPT6, KOWx-4 selected particles (Map D)
Method: EM (single particle) / Resolution: 3.2 Å

EMDB-0034:
Structure of activated transcription complex Pol II-DSIF-PAF-SPT6, SPT6 core selected particles (Map E)
Method: EM (single particle) / Resolution: 3.28 Å

EMDB-0035:
Structure of activated transcription complex Pol II-DSIF-PAF-SPT6, selected SPT6 tSH2 particles (Map F)
Method: EM (single particle) / Resolution: 3.49 Å

EMDB-0036:
Structure of activated transcription complex Pol II-DSIF-PAF-SPT6, upstream DNA selected particles (Map G)
Method: EM (single particle) / Resolution: 3.1 Å

EMDB-0037:
Structure of activated transcription complex Pol II-DSIF-PAF-SPT6, CTR9 selected particles (Map H)
Method: EM (single particle) / Resolution: 3.34 Å

PDB-6gme:
Structure of H. sapiens SPT6 tandem SH2 domain
Method: X-RAY DIFFRACTION / Resolution: 1.802 Å

Chemicals

ChemComp-HOH:
WATER / Water

ChemComp-ZN:
Unknown entry

ChemComp-MG:
Unknown entry

Source
  • sus scrofa (pig)
  • homo sapiens (human)
  • synthetic construct (others)
  • Pig (pig)
KeywordsTRANSCRIPTION / SH2 domain / Pol II interaction / Kinase target / Transcription factor / DNA / RNA Polymerase / DSIF / PAF1c / SPT6

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