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9J0N

Paused elongation complex of mammalian RNA polymerase II with nucleosome (PEC2-nuc)

Summary for 9J0N
Entry DOI10.2210/pdb9j0n/pdb
EMDB information61058
DescriptorDNA-directed RNA polymerase subunit, Transcription elongation factor SPT5, Non-template DNA (157-MER), ... (28 entities in total)
Functional Keywordsrna polymerase, nelf, nucleosome, dna, rna, tfiis, transcription
Biological sourceHomo sapiens (human)
More
Total number of polymer chains30
Total formula weight1171165.15
Authors
Naganuma, M.,Kujirai, T.,Ehara, H.,Uejima, T.,Ito, T.,Goto, M.,Aoki, M.,Henmi, M.,Miyamoto-Kohno, S.,Shirouzu, M.,Kurumizaka, H.,Sekine, S. (deposition date: 2024-08-02, release date: 2025-02-05, Last modification date: 2025-03-19)
Primary citationNaganuma, M.,Kujirai, T.,Ehara, H.,Uejima, T.,Ito, T.,Goto, M.,Aoki, M.,Henmi, M.,Miyamoto-Kohno, S.,Shirouzu, M.,Kurumizaka, H.,Sekine, S.I.
Structural insights into promoter-proximal pausing of RNA polymerase II at +1 nucleosome.
Sci Adv, 11:eadu0577-eadu0577, 2025
Cited by
PubMed Abstract: The metazoan transcription elongation complex (EC) of RNA polymerase II (RNAPII) generally stalls between the transcription start site and the first (+1) nucleosome. This promoter-proximal pausing involves negative elongation factor (NELF), 5,6-dichloro-1-β-d-ribobenzimidazole sensitivity-inducing factor (DSIF), and transcription elongation factor IIS (TFIIS) and is critical for subsequent productive transcription elongation. However, the detailed pausing mechanism and the involvement of the +1 nucleosome remain enigmatic. Here, we report cryo-electron microscopy structures of ECs stalled on nucleosomal DNA. In the absence of TFIIS, the EC is backtracked/arrested due to conflicts between NELF and the nucleosome. We identified two alternative binding modes of NELF, one of which reveals a critical contact with the downstream DNA through the conserved NELF-E basic helix. Upon binding with TFIIS, the EC progressed to the nucleosome to establish a paused EC with a partially unwrapped nucleosome. This paused EC strongly restricts EC progression further downstream. These structures illuminate the mechanism of RNAPII pausing/stalling at the +1 nucleosome.
PubMed: 40043114
DOI: 10.1126/sciadv.adu0577
PDB entries with the same primary citation
Experimental method
ELECTRON MICROSCOPY (3.4 Å)
Structure validation

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