Transcription-coupled nucleotide excision repair of a gene transcribed by bacteriophage T7 RNA polymerase in Escherichia coli.

DNA Repair
Ann K Ganesan, Philip C Hanawalt

Abstract

Transcription-coupled nucleotide excision repair (TC-NER) removes certain kinds of lesions from the transcribed strand of expressed genes. The signal for TC-NER is thought to be RNA polymerase stalled at a lesion in the DNA template. In Escherichia coli, the stalled polymerase is dissociated from the lesion by the transcription repair coupling factor (Mfd protein), which also recruits excision repair proteins to the site resulting in efficient removal of the lesion. TC-NER has been documented in cells from a variety of organisms ranging from bacteria to humans. In each case, the RNA polymerase involved has been a multimeric protein complex. To ascertain whether a gene transcribed by the monomeric RNA polymerase of bacteriophage T7 could be repaired by TC-NER, we constructed strains of E. coli in which the chromosomal lacZ gene is controlled by a T7 promoter. In the absence of T7 RNA polymerase, little or no beta-galactosidase is produced, indicating that the E. coli RNA polymerase does not transcribe lacZ efficiently, if at all, in these strains. By introducing a plasmid (pAR1219) carrying the T7 gene 1 under control of the E. coli lac UV5 promoter into these strains, we obtained derivatives in which the level of T7 RNA polymer...Continue Reading

Citations

Oct 23, 2012·Trends in Biochemical Sciences·Alexandra M DeaconescuNikolaus Grigorieff
Jun 26, 2012·Biochimica Et Biophysica Acta·Susan D Cline
Oct 11, 2012·Biochimica Et Biophysica Acta·Hélène Gaillard, Andrés Aguilera
Jan 18, 2015·Current Opinion in Microbiology·Venu Kamarthapu, Evgeny Nudler
Nov 20, 2016·Photochemistry and Photobiology·Christopher P Selby
Apr 27, 2011·FEMS Microbiology Reviews·Silvia AyoraJuan C Alonso
Mar 22, 2020·Nature Communications·Harshad GhodkeAntoine M van Oijen

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