7KED

RNA polymerase II elongation complex with unnatural base dTPT3

7KED の概要

• エントリーDOI: 10.2210/pdb7ked/pdb
• 分子名称: DNA-directed RNA polymerase II subunit RPB1, DNA-directed RNA polymerases I, II, and III subunit RPABC4, RNA, ... (15 entities in total)
• 機能のキーワード: rna polymerase ii, elongation complex, unnatural base, tpt3, nam, transcription, transcription-dna-rna complex, transcription/dna/rna
• 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: 13
• 化学式量合計: 487253.63

構造登録者

Oh, J.,Wang, W.,Wang, D. (登録日: 2020-10-10, 公開日: 2021-06-23, 最終更新日: 2023-10-18)

主引用文献

Oh, J.,Shin, J.,Unarta, I.C.,Wang, W.,Feldman, A.W.,Karadeema, R.J.,Xu, L.,Xu, J.,Chong, J.,Krishnamurthy, R.,Huang, X.,Romesberg, F.E.,Wang, D.
Transcriptional processing of an unnatural base pair by eukaryotic RNA polymerase II.
Nat.Chem.Biol., 17:906-914, 2021

PubMed Abstract: The development of unnatural base pairs (UBPs) has greatly increased the information storage capacity of DNA, allowing for transcription of unnatural RNA by the heterologously expressed T7 RNA polymerase (RNAP) in Escherichia coli. However, little is known about how UBPs are transcribed by cellular RNA polymerases. Here, we investigated how synthetic unnatural nucleotides, NaM and TPT3, are recognized by eukaryotic RNA polymerase II (Pol II) and found that Pol II is able to selectively recognize UBPs with high fidelity when dTPT3 is in the template strand and rNaMTP acts as the nucleotide substrate. Our structural analysis and molecular dynamics simulation provide structural insights into transcriptional processing of UBPs in a stepwise manner. Intriguingly, we identified a novel 3'-RNA binding site after rNaM addition, termed the swing state. These results may pave the way for future studies in the design of transcription and translation strategies in higher organisms with expanded genetic codes.

PubMed: 34140682
DOI: 10.1038/s41589-021-00817-3

実験手法

X-RAY DIFFRACTION (3.6 Å)