replication fork reversal after replication–transcription collision复制叉replication-transcription碰撞后逆转.pdfVIP

Replication Fork Reversal after Replication–Transcription Collision 1,2 ´ 1,2. 1,2.¤ ´ ´ 1,2 Anne L. De Septenville , Stephane Duigou , Hasna Boubakri , Benedicte Michel * ´ ´ ´ ´ 1 CNRS, Centre de Genetique Moleculaire, UPR3404, Gif-sur-Yvette, France, 2 Universite Paris-Sud, Orsay, France Abstract Replication fork arrest is a recognized source of genetic instability, and transcription is one of the most prominent causes of replication impediment. We analyze here the requirement for recombination proteins in Escherichia coli when replication– transcription head-on collisions are induced at a specific site by the inversion of a highly expressed ribosomal operon (rrn). RecBC is the only recombination protein required for cell viability under these conditions of increased replication- transcription collisions. In its absence, fork breakage occurs at the site of collision, and the resulting linear DNA is not repaired and is slowly degraded by the RecJ exonuclease. Lethal fork breakage is also observed in cells that lack RecA and RecD, i.e. when both homologous recombination and the potent exonuclease V activity of the RecBCD complex are inactivated, with a slow degradation of the resulting linear DNA by the combined action of the RecBC helicase and the RecJ exonuclease. The sizes of the major linear fragments indicate that DNA degradation is slowed down by the encounter with another rrn operon. The amount of linear DNA decreases nearly two-fold when the Holliday junction resolvase RuvABC is inactivated in recB, as well as in rec