5OT2

RNA polymerase II elongation complex in the presence of 3d-Napht-A

5OT2 の概要

• エントリーDOI: 10.2210/pdb5ot2/pdb
• 分子名称: DNA-directed RNA polymerase II subunit RPB1, DNA-directed RNA polymerases I, II, and III subunit RPABC5, DNA-directed RNA polymerase II subunit RPB11, ... (18 entities in total)
• 機能のキーワード: inhibitor, complex, polymerase, transcription
• 由来する生物種: Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (Baker's yeast); 詳細
• タンパク質・核酸の鎖数: 15
• 化学式量合計: 530696.01

構造登録者

Malvezzi, S.,Farnung, L.,Aloisi, C.,Angelov, T.,Cramer, P.,Sturla, S.J. (登録日: 2017-08-19, 公開日: 2017-11-01, 最終更新日: 2025-07-30)

主引用文献

Malvezzi, S.,Farnung, L.,Aloisi, C.M.N.,Angelov, T.,Cramer, P.,Sturla, S.J.
Mechanism of RNA polymerase II stalling by DNA alkylation.
Proc. Natl. Acad. Sci. U.S.A., 114:12172-12177, 2017

PubMed Abstract: Several anticancer agents that form DNA adducts in the minor groove interfere with DNA replication and transcription to induce apoptosis. Therapeutic resistance can occur, however, when cells are proficient in the removal of drug-induced damage. Acylfulvenes are a class of experimental anticancer agents with a unique repair profile suggesting their capacity to stall RNA polymerase (Pol) II and trigger transcription-coupled nucleotide excision repair. Here we show how different forms of DNA alkylation impair transcription by RNA Pol II in cells and with the isolated enzyme and unravel a mode of RNA Pol II stalling that is due to alkylation of DNA in the minor groove. We incorporated a model for acylfulvene adducts, the stable 3-deaza-3-methoxynaphtylethyl-adenosine analog (3d-Napht-A), and smaller 3-deaza-adenosine analogs, into DNA oligonucleotides to assess RNA Pol II transcription elongation in vitro. RNA Pol II was strongly blocked by a 3d-Napht-A analog but bypassed smaller analogs. Crystal structure analysis revealed that a DNA base containing 3d-Napht-A can occupy the +1 templating position and impair closing of the trigger loop in the Pol II active center and polymerase translocation into the next template position. These results show how RNA Pol II copes with minor-groove DNA alkylation and establishes a mechanism for drug resistance.

PubMed: 29087308
DOI: 10.1073/pnas.1706592114

実験手法

X-RAY DIFFRACTION (3.2 Å)