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- W2912507699 abstract "The process of bacterial RNA polymerase (RNAP) escaping the promoter region during transcription initiation can often be rate limiting, making kinetic regulation of escape a common strategy employed to control the rates of transcription. Using a pre-steady state ensemble fluorescence assay, we present the first in-depth kinetic analysis of initial nucleotide incorporation and promoter escape for the Mycobacterium tuberculosis (Mtb) RNAP.We show that in relation to E. coli RNAP, Mtb displays slower initial nucleotide incorporation but faster overall promoter escape kinetics on the MtbrrnAP3 promoter. Furthermore, in the context of the essential transcription factors CarD and RbpA, Mtb RNAP promoter escape is delayed, albeit through different mechanisms. We also see that these factors increase the inactive fraction, i.e. complexes that are unable to escape the promoter. Given that CarD and RbpA affect multiple kinetic intermediates during transcription initiation, we modelled hypothetical sets of rate constants derived from the initiation kinetics to suggest that CarD and RbpA can serve to both activate and repress transcription in a promoter-dependent context. For instance, on the rrnAP3 promoter, where DNA-opening is rate limiting, both CarD and RbpA activate transcription, whereas on promoters that are rate-limited at escape, the prediction is that these factors lead to transcriptional repression." @default.
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- W2912507699 date "2019-02-01" @default.
- W2912507699 modified "2023-09-26" @default.
- W2912507699 title "Regulation of Mycobacterial RNA Polymerase Promoter Escape Kinetics by Transcription Factors Card and RbpA" @default.
- W2912507699 doi "https://doi.org/10.1016/j.bpj.2018.11.1160" @default.
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