Transcriptional processing of an unnatural base pair by eukaryotic RNA polymerase II.

Autor: Oh J; Division of Pharmaceutical Sciences, Skaggs School of Pharmacy & Pharmaceutical Sciences, University of California, San Diego, La Jolla, CA, USA., Shin J; Division of Pharmaceutical Sciences, Skaggs School of Pharmacy & Pharmaceutical Sciences, University of California, San Diego, La Jolla, CA, USA.; Department of Molecular Microbiology, Center for Advanced Laboratory Medicine, University of California, San Diego, La Jolla, CA, USA., Unarta IC; Department of Chemistry, The Hong Kong University of Science and Technology, Kowloon, Hong Kong., Wang W; Division of Pharmaceutical Sciences, Skaggs School of Pharmacy & Pharmaceutical Sciences, University of California, San Diego, La Jolla, CA, USA.; Advanced Medical Research Institute, Shandong University, Jinan, China., Feldman AW; Department of Chemistry, The Scripps Research Institute, La Jolla, CA, USA., Karadeema RJ; Department of Chemistry, The Scripps Research Institute, La Jolla, CA, USA., Xu L; Division of Pharmaceutical Sciences, Skaggs School of Pharmacy & Pharmaceutical Sciences, University of California, San Diego, La Jolla, CA, USA.; Department of Chemistry, Sun Yat-Sen University, Guangzhou, China., Xu J; Division of Pharmaceutical Sciences, Skaggs School of Pharmacy & Pharmaceutical Sciences, University of California, San Diego, La Jolla, CA, USA., Chong J; Division of Pharmaceutical Sciences, Skaggs School of Pharmacy & Pharmaceutical Sciences, University of California, San Diego, La Jolla, CA, USA., Krishnamurthy R; Department of Chemistry, The Scripps Research Institute, La Jolla, CA, USA., Huang X; Department of Chemistry, The Hong Kong University of Science and Technology, Kowloon, Hong Kong., Romesberg FE; Synthorx, La Jolla, CA, USA., Wang D; Division of Pharmaceutical Sciences, Skaggs School of Pharmacy & Pharmaceutical Sciences, University of California, San Diego, La Jolla, CA, USA. dongwang@ucsd.edu.; Department of Cellular and Molecular Medicine, University of California, San Diego, La Jolla, CA, USA. dongwang@ucsd.edu.; Department of Chemistry and Biochemistry, University of California, San Diego, La Jolla, CA, USA. dongwang@ucsd.edu.
Jazyk: angličtina
Zdroj: Nature chemical biology [Nat Chem Biol] 2021 Aug; Vol. 17 (8), pp. 906-914. Date of Electronic Publication: 2021 Jun 17.
DOI: 10.1038/s41589-021-00817-3
Abstrakt: 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.
(© 2021. The Author(s), under exclusive licence to Springer Nature America, Inc.)
Databáze: MEDLINE
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