Transcription-Coupled Repair: From Cells to Single Molecules and Back Again.

Transcription-coupled repair is mediated by the Mfd protein. TCR is defined as the preferential repair of DNA lesions in the transcribed strand of actively transcribed genes, and is opposed to the strand-aspecific global genome repair. The Mfd protein mediates TCR by binding to and displacing RNA polymerase, which is stalled at a DNA lesion on the transcribed strand of DNA, then recruiting UvrA and UvrB. The repair cascade results in the recruitment of, and DNA excision by, UvrC; removal of the damage-bearing oligonucleotide by UvrD; "filling-in" of the DNA by DNA polymerase; and sealing of the strands by DNA ligase. The gene required for Mfd was originally identified as a gene needed for the "mutation frequency decline" phenotype in which the repair of certain UV-induced lesions in the transcribed strand of tRNA genes is increased when cells are forced to delay replication immediately following UV exposure. This review will focus on the genetics that led to the discovery of the Mfd gene; summarize the subsequent biochemical, structural and single-molecule interrogations of the Mfd protein; and explore the more recent findings of Mfd in mutagenesis.