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-Structure paper
| タイトル | Structural and kinetic insights into tRNA promoter engagement by yeast general transcription factor TFIIIC. |
|---|---|
| ジャーナル・号・ページ | Nucleic Acids Res, Vol. 53, Issue 1, Year 2025 |
| 掲載日 | 2025年1月7日 |
著者 | Wolfram Seifert-Dávila / Anastasiia Chaban / Florence Baudin / Mathias Girbig / Luis Hauptmann / Thomas Hoffmann / Olivier Duss / Sebastian Eustermann / Christoph W Müller / ![]() |
| PubMed 要旨 | Transcription of transfer RNA (tRNA) genes by RNA polymerase (Pol) III requires the general transcription factor IIIC (TFIIIC), which recognizes intragenic A-box and B-box DNA motifs of type II ...Transcription of transfer RNA (tRNA) genes by RNA polymerase (Pol) III requires the general transcription factor IIIC (TFIIIC), which recognizes intragenic A-box and B-box DNA motifs of type II gene promoters. However, the underlying mechanism has remained elusive, in part due to missing structural information for A-box recognition. In this study, we use single-particle cryogenic electron microscopy (cryo-EM) and single-molecule fluorescence resonance energy transfer (smFRET) to reveal structural and real-time kinetic insights into how the 520-kDa yeast TFIIIC complex engages A-box and B-box DNA motifs in the context of a tRNA gene promoter. Cryo-EM structures of τA and τB subcomplexes bound to the A-box and B-box were obtained at 3.7 and 2.5 Å resolution, respectively, while cryo-EM single-particle mapping determined the specific distance and relative orientation of the τA and τB subcomplexes revealing a fully engaged state of TFIIIC. smFRET experiments show that overall recruitment and residence times of TFIIIC on a tRNA gene are primarily governed by B-box recognition, while footprinting experiments suggest a key role of τA and the A-box in TFIIIB and Pol III recruitment following TFIIIC recognition of type II promoters. |
リンク | Nucleic Acids Res / PubMed:39657784 / PubMed Central |
| 手法 | EM (単粒子) |
| 解像度 | 2.46 - 3.7 Å |
| 構造データ | EMDB-51228, PDB-9gc3: EMDB-51231, PDB-9gck: |
| 由来 |
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キーワード | DNA BINDING PROTEIN / Base-specific contacts / B-box / tRNA gene / A-box |
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