Benefit of transcription-coupled nucleotide excision repair for gene expression in u.v.-damaged Escherichia coli.
Expression of the lactose operon upon induction by IPTG was studied with Escherichia coli B/r and K-12 strains as a function of exposure to ultraviolet light. Patterns of expression inactivation were compared in cells with wild-type UvrABC nucleotide excision repair, with transcription-coupled excision repair (TCR) specifically defective because of a defect at mfd, or with excision repair (ER) and TCR eliminated by defects at uvrA or uvrC. Sets of inactivation patterns were also determined for cells expressing the lactose operon via the "UV5' promoter, an alternative to the wild-type promoter that eliminates dependence of expression on negative DNA supercoiling. The results demonstrated a major contribution by TCR to successful gene expression. Gene expression was more sensitive to u.v. inactivation when TCR was defective and similarly more sensitive when both ER and TCR were defective. Thus, TCR may be the only means of repairing transcription-blocking damage at active genes. Contrasting results with wild-type and UV5 promoters suggested that relaxed supercoiling might accompany repair and reduce expression even though a template lesion is removed. A test of mismatch repair defects on ultraviolet inactivation of gene expression found only limited interference with TCR as it benefits gene expression.
Li
B H
BH
Department of Microbiology and Immunology, Indiana University School of Medicine, Indianapolis 46202, USA.
Bockrath
R
R
D000251:Adenosine Triphosphatases / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics; D004260:DNA Repair / Q000235:genetics*; D004278:DNA, Superhelical / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics; D004706:Endodeoxyribonucleases / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics*; D029968:Escherichia coli Proteins; D015964:Gene Expression Regulation, Bacterial / Q000528:radiation effects*; D005798:Genes, Bacterial; D007763:Lac Operon / Q000528:radiation effects*; D011401:Promoter Regions, Genetic / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics; D014158:Transcription, Genetic; D014466:Ultraviolet Rays / Q000009:adverse effects*
D001426:Bacterial Proteins; D004278:DNA, Superhelical; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; D014157:Transcription Factors; C072103:transcription repair coupling factor protein, Bacteria; D004706:Endodeoxyribonucleases; C066567:UvrC protein, E coli; C036385:endodeoxyribonuclease uvrABC; C068631:UvrA protein, E coli; D000251:Adenosine Triphosphatases