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Structure paper

TitleStructural basis of nucleosome transcription mediated by Chd1 and FACT.
Journal, issue, pagesNat Struct Mol Biol, Vol. 28, Issue 4, Page 382-387, Year 2021
Publish dateApr 12, 2021
AuthorsLucas Farnung / Moritz Ochmann / Maik Engeholm / Patrick Cramer /
PubMed AbstractEfficient transcription of RNA polymerase II (Pol II) through nucleosomes requires the help of various factors. Here we show biochemically that Pol II transcription through a nucleosome is ...Efficient transcription of RNA polymerase II (Pol II) through nucleosomes requires the help of various factors. Here we show biochemically that Pol II transcription through a nucleosome is facilitated by the chromatin remodeler Chd1 and the histone chaperone FACT when the elongation factors Spt4/5 and TFIIS are present. We report cryo-EM structures of transcribing Saccharomyces cerevisiae Pol II-Spt4/5-nucleosome complexes with bound Chd1 or FACT. In the first structure, Pol II transcription exposes the proximal histone H2A-H2B dimer that is bound by Spt5. Pol II has also released the inhibitory DNA-binding region of Chd1 that is poised to pump DNA toward Pol II. In the second structure, Pol II has generated a partially unraveled nucleosome that binds FACT, which excludes Chd1 and Spt5. These results suggest that Pol II progression through a nucleosome activates Chd1, enables FACT binding and eventually triggers transfer of FACT together with histones to upstream DNA.
External linksNat Struct Mol Biol / PubMed:33846633 / PubMed Central
MethodsEM (single particle)
Resolution2.7 - 3.5 Å
Structure data

12449

7nkx

EMDB-12449, PDB-7nkx:
RNA polymerase II-Spt4/5-nucleosome-Chd1 structure
Method: EM (single particle) / Resolution: 2.9 Å

12450

7nky

EMDB-12450, PDB-7nky:
RNA Polymerase II-Spt4/5-nucleosome-FACT structure
Method: EM (single particle) / Resolution: 3.2 Å

EMDB-12666:
RNA polymerase II-Spt4/5-nucleosome-Chd1 structure - Map A
Method: EM (single particle) / Resolution: 2.9 Å

EMDB-12667:
RNA polymerase II-Spt4/5-nucleosome-Chd1 structure - Map B
Method: EM (single particle) / Resolution: 2.7 Å

EMDB-12668:
RNA polymerase II-Spt4/5-nucleosome-Chd1 structure - Map C
Method: EM (single particle) / Resolution: 3.1 Å

EMDB-12669:
RNA polymerase II-Spt4/5-nucleosome-FACT structure - Map 1
Method: EM (single particle) / Resolution: 3.1 Å

EMDB-12670:
RNA polymerase II-Spt4/5-nucleosome-FACT structure - Map 2
Method: EM (single particle) / Resolution: 3.0 Å

EMDB-12671:
RNA polymerase II-Spt4/5-nucleosome-FACT structure - Map 3
Method: EM (single particle) / Resolution: 3.5 Å

Chemicals

ChemComp-ZN:
Unknown entry

ChemComp-MG:
Unknown entry

ChemComp-ADP:
ADENOSINE-5'-DIPHOSPHATE / ADP, energy-carrying molecule*YM

ChemComp-BEF:
BERYLLIUM TRIFLUORIDE ION

Source
  • saccharomyces cerevisiae (brewer's yeast)
  • xenopus laevis (African clawed frog)
  • saccharomyces cerevisiae (strain atcc 204508 / s288c) (yeast)
  • synthetic construct (others)
  • synthetic rna (others)
KeywordsTRANSCRIPTION / chromatin remodelling / nucleosome / chromatin / elongation / histone chaperone

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