T7 RNA polymerase transcription with 5-position modified UTP derivatives

Journal of the American Chemical Society
Jonathan D VaughtBruce E Eaton

Abstract

Seven UTP derivatives modified at the 5-position through an amide linkage were tested as substrates for T7 RNA polymerase (T7 RNAP) transcription. All UTP derivatives gave good yields of full-length transcript even from DNA templates that showed a significant number of abortive transcripts using unmodified UTP. A kinetic assay to determine the relative K(m) and V(max) for T7 RNAP transcription gave surprisingly similar values for UTP and the 5-position hydrophobic modifications phenyl, 4-pyridyl, 2-pyridyl, indolyl, and isobutyl. The 5-position modifications imidazole and amino, which could both be positively charged, gave K(m) values significantly higher than UTP. All seven UTP derivatives gave relative V(max) values similar to UTP, indicating that insertion of these modified bases into the transcript did not impede its elongation.

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Citations

Apr 29, 2011·Analytical Chemistry·Anton B IliukW Andy Tao
Mar 6, 2010·Journal of the American Chemical Society·Jonathan D VaughtBruce E Eaton
Jul 2, 2010·Nature Reviews. Drug Discovery·Anthony D KeefeAndrew Ellington
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Jan 6, 2017·Journal of the American Chemical Society·Chun Pong TamJack W Szostak
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