4.6 Review

Molecular Mechanisms of Transcription-Coupled Repair

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Summary: Elongation by RNA polymerase can be dynamically modulated by accessory factors, and the transcription-repair coupling factor TRCF can choose to either rescue or terminate the paused/stalled RNAPs. The mechanism for TRCF's choice remains unclear. Using single-molecule assays with Escherichia coli as a model, it was found that nucleotide-bound Mfd, a bacterial TRCF, can convert the elongation complex (EC) into a catalytically poised state, allowing the EC to restart transcription. After a prolonged residence in this state, ATP hydrolysis by Mfd leads to a remodeling of the EC and loss of the RNA transcript. Biophysical studies also revealed that the motor domain of Mfd can bind and partially melt DNA with a template strand overhang.

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Summary: Transcription-coupled repair (TCR), a sub-pathway of nucleotide excision repair (NER), removes lesions from the template-strand that stall RNA polymerase elongation complexes. Cryo-electron microscopy was used to visualize Mfd engaging with a stalled EC and dislodging the RNAP, showing how Mfd discriminates between paused and stalled ECs and initiates TCR through a complex mechanism.
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Summary: DNA lesions are a major obstacle during gene transcription by RNA polymerase II enzymes, but can be eliminated by the transcription-coupled DNA repair pathway. Recent advances suggest that RNAPII ubiquitylation activates TCR complex assembly during repair and promotes processing and degradation of RNAPII to prevent prolonged stalling, making the fate of stalled RNAPII a crucial link between TCR and associated human diseases.

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