Dataset Information


Functional interplay between NTP leaving group and base pair recognition during RNA polymerase II nucleotide incorporation revealed by methylene substitution.

ABSTRACT: RNA polymerase II (pol II) utilizes a complex interaction network to select and incorporate correct nucleoside triphosphate (NTP) substrates with high efficiency and fidelity. Our previous 'synthetic nucleic acid substitution' strategy has been successfully applied in dissecting the function of nucleic acid moieties in pol II transcription. However, how the triphosphate moiety of substrate influences the rate of P-O bond cleavage and formation during nucleotide incorporation is still unclear. Here, by employing β,γ-bridging atom-'substituted' NTPs, we elucidate how the methylene substitution in the pyrophosphate leaving group affects cognate and non-cognate nucleotide incorporation. Intriguingly, the effect of the β,γ-methylene substitution on the non-cognate UTP/dT scaffold (∼3-fold decrease in kpol) is significantly different from that of the cognate ATP/dT scaffold (∼130-fold decrease in kpol). Removal of the wobble hydrogen bonds in U:dT recovers a strong response to methylene substitution of UTP. Our kinetic and modeling studies are consistent with a unique altered transition state for bond formation and cleavage for UTP/dT incorporation compared with ATP/dT incorporation. Collectively, our data reveals the functional interplay between NTP triphosphate moiety and base pair hydrogen bonding recognition during nucleotide incorporation.


PROVIDER: S-EPMC4857003 | BioStudies | 2016-01-01


REPOSITORIES: biostudies

Similar Datasets

1000-01-01 | S-EPMC4068585 | BioStudies
1000-01-01 | S-EPMC6181931 | BioStudies
2014-01-01 | S-EPMC4119285 | BioStudies
1998-01-01 | S-EPMC1219323 | BioStudies
2016-01-01 | S-EPMC5081176 | BioStudies
2017-01-01 | S-EPMC5522186 | BioStudies
2018-01-01 | S-EPMC6039227 | BioStudies
2011-01-01 | S-EPMC3243610 | BioStudies
2007-01-01 | S-EPMC1976215 | BioStudies
2016-01-01 | S-EPMC5749642 | BioStudies