title: Response of the bacteriophage T4 replisome to noncoding lesions and regression of a stalled replication fork.
abstract: DNA is constantly damaged by endogenous and exogenous agents. The resulting DNA lesions have the potential to halt the progression of the replisome, possibly leading to replication fork collapse. Here, we examine the effect of a noncoding DNA lesion in either leading strand template or lagging strand template on the bacteriophage T4 replisome. A damaged base in the lagging strand template does not affect the progression of the replication fork. Instead, the stalled lagging strand polymerase recycles from the lesion and initiates the synthesis of a new Okazaki fragment upstream of the damaged base. In contrast, when the replisome encounters a blocking lesion in the leading strand template, the replication fork only travels approximately 1 kb beyond the point of the DNA lesion before complete replication fork collapse. The primosome and the lagging strand polymerase remain active during this period, and an Okazaki fragment is synthesized beyond the point of the leading strand lesion. There is no evidence for a new priming event on the leading strand template. Instead, the DNA structure that is produced by the stalled replication fork is a substrate for the DNA repair helicase UvsW. UvsW catalyzes the regression of a stalled replication fork into a "chicken-foot" structure that has been postulated to be an intermediate in an error-free lesion bypass pathway.
authors.affiliation: Department of Biochemistry, 4112 Molecular Biology Building, Iowa State University, Ames, IA 50011, USA. swn@iastate.edu
authors.forename: Scott W
authors.initials: SW
authors.lastname: Nelson
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Benkovic
mesh_terms: D017122:Bacteriophage T4 / Q000235:genetics / Q000502:physiology*; D004247:DNA; D004265:DNA Helicases / Q000502:physiology; D004261:DNA Replication; D004587:Electrophoresis, Agar Gel; D014764:Viral Proteins / Q000502:physiology
chemical_list: C022262:Okazaki fragments; D014764:Viral Proteins; D004247:DNA; D004265:DNA Helicases; C107414:UvsW protein, Bacteriophage T4