National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)
GM102362
米国
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)
GM148476
米国
National Institutes of Health/National Institute Of Allergy and Infectious Diseases (NIH/NIAID)
U54 AI170855
米国
National Science Foundation (NSF, United States)
MCB-2048095
米国
National Institutes of Health/National Cancer Institute (NIH/NCI)
1R21CA251043-01A1
米国
National Science Foundation (NSF, United States)
MCB- 2123995
米国
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)
U24 GM129541
米国
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)
S10 OD032467
米国
引用
ジャーナル: Nat Commun / 年: 2023 タイトル: A unified Watson-Crick geometry drives transcription of six-letter expanded DNA alphabets by E. coli RNA polymerase. 著者: Juntaek Oh / Zelin Shan / Shuichi Hoshika / Jun Xu / Jenny Chong / Steven A Benner / Dmitry Lyumkis / Dong Wang / 要旨: Artificially Expanded Genetic Information Systems (AEGIS) add independently replicable unnatural nucleotide pairs to the natural G:C and A:T/U pairs found in native DNA, joining the unnatural pairs ...Artificially Expanded Genetic Information Systems (AEGIS) add independently replicable unnatural nucleotide pairs to the natural G:C and A:T/U pairs found in native DNA, joining the unnatural pairs through alternative modes of hydrogen bonding. Whether and how AEGIS pairs are recognized and processed by multi-subunit cellular RNA polymerases (RNAPs) remains unknown. Here, we show that E. coli RNAP selectively recognizes unnatural nucleobases in a six-letter expanded genetic system. High-resolution cryo-EM structures of three RNAP elongation complexes containing template-substrate UBPs reveal the shared principles behind the recognition of AEGIS and natural base pairs. In these structures, RNAPs are captured in an active state, poised to perform the chemistry step. At this point, the unnatural base pair adopts a Watson-Crick geometry, and the trigger loop is folded into an active conformation, indicating that the mechanistic principles underlying recognition and incorporation of natural base pairs also apply to AEGIS unnatural base pairs. These data validate the design philosophy of AEGIS unnatural basepairs. Further, we provide structural evidence supporting a long-standing hypothesis that pair mismatch during transcription occurs via tautomerization. Together, our work highlights the importance of Watson-Crick complementarity underlying the design principles of AEGIS base pair recognition.
超分子 #1: E. coli RNAP transcription elongation complex bound the unnatural...
超分子
名称: E. coli RNAP transcription elongation complex bound the unnatural dS-BTP base pair in the active site タイプ: complex / ID: 1 / 親要素: 0 / 含まれる分子: #1-#7