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-Structure paper
| タイトル | Transcription start site scanning requires the fungi-specific hydrophobic loop of Tfb3. |
|---|---|
| ジャーナル・号・ページ | Nucleic Acids Res, Vol. 52, Issue 19, Page 11602-11611, Year 2024 |
| 掲載日 | 2024年10月28日 |
著者 | Chun Yang / Pratik Basnet / Samah Sharmin / Hui Shen / Craig D Kaplan / Kenji Murakami / ![]() |
| PubMed 要旨 | RNA polymerase II (pol II) initiates transcription from transcription start sites (TSSs) located ∼30-35 bp downstream of the TATA box in metazoans, whereas in the yeast Saccharomyces cerevisiae, ...RNA polymerase II (pol II) initiates transcription from transcription start sites (TSSs) located ∼30-35 bp downstream of the TATA box in metazoans, whereas in the yeast Saccharomyces cerevisiae, pol II scans further downstream TSSs located ∼40-120 bp downstream of the TATA box. Previously, we found that removal of the kinase module TFIIK (Kin28-Ccl1-Tfb3) from TFIIH shifts the TSS in a yeast in vitro system upstream to the location observed in metazoans and that addition of recombinant Tfb3 back to TFIIH-ΔTFIIK restores the downstream TSS usage. Here, we report that this biochemical activity of yeast TFIIK in TSS scanning is attributable to the Tfb3 RING domain at the interface with pol II in the pre-initiation complex (PIC): especially, swapping Tfb3 Pro51-a residue conserved among all fungi-with Ala or Ser as in MAT1, the metazoan homolog of Tfb3, confers an upstream TSS shift in vitro in a similar manner to the removal of TFIIK. Yeast genetic analysis suggests that both Pro51 and Arg64 of Tfb3 are required to maintain the stability of the Tfb3-pol II interface in the PIC. Cryo-electron microscopy analysis of a yeast PIC lacking TFIIK reveals considerable variability in the orientation of TFIIH, which impairs TSS scanning after promoter opening. |
リンク | Nucleic Acids Res / PubMed:39287137 / PubMed Central |
| 手法 | EM (単粒子) |
| 解像度 | 3.7 - 3.8 Å |
| 構造データ | EMDB-42437, PDB-8uoq: EMDB-42438, PDB-8uot: |
| 化合物 | ![]() ChemComp-ZN: ![]() ChemComp-MG: ![]() ChemComp-SF4: |
| 由来 |
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キーワード | TRANSCRIPTION / pol II / Transcription. POL II |
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