The helicases DinG, Rep and UvrD cooperate to promote replication across transcription units in vivo.
How living cells deal with head-on collisions of the replication and transcription complexes has been debated for a long time. Even in the widely studied model bacteria Escherichia coli, the enzymes that take care of such collisions are still unknown. We report here that in vivo, the DinG, Rep and UvrD helicases are essential for efficient replication across highly transcribed regions. We show that when rRNA operons (rrn) are inverted to face replication, the viability of the dinG mutant is affected and over-expression of RNase H rescues the growth defect, showing that DinG acts in vivo to remove R-loops. In addition, DinG, Rep and UvrD exert a common function, which requires the presence of two of these three helicases. After replication blockage by an inverted rrn, Rep in conjunction with DinG or UvrD removes RNA polymerase, a task that is fulfilled in its absence by the SOS-induced DinG and UvrD helicases. Finally, Rep and UvrD also act at inverted sequences other than rrn, and promote replication through highly transcribed regions in wild-type E. coli.
Boubakri
Hasna
H
CNRS, Centre de Génétique Moléculaire, FRE 3144, Gif-sur-Yvette, France.
de Septenville
Anne Langlois
AL
Viguera
Enrique
E
Michel
Bénédicte
B
D004265:DNA Helicases / Q000235:genetics / Q000378:metabolism*; D004261:DNA Replication / Q000235:genetics / Q000502:physiology*; D004269:DNA, Bacterial / Q000096:biosynthesis; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D048168:Escherichia coli K12 / Q000166:cytology / Q000235:genetics / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism*; D005798:Genes, Bacterial; D008954:Models, Biological; D009154:Mutation; D057345:Sequence Inversion; D014158:Transcription, Genetic; D012407:rRNA Operon
D004269:DNA, Bacterial; D029968:Escherichia coli Proteins; C074432:dinG protein, E coli; C506744:rep protein, E coli; D012321:DNA-Directed RNA Polymerases; C506746:UvrD protein, E coli; D004265:DNA Helicases