title: Dynamics of leading-strand lesion skipping by the replisome.
abstract: The E. coli replisome stalls transiently when it encounters a lesion in the leading-strand template, skipping over the damage by reinitiating replication at a new primer synthesized downstream by the primase. We report here that template unwinding and lagging-strand synthesis continue downstream of the lesion at a reduced rate after replisome stalling, that one replisome is capable of skipping multiple lesions, and that the rate-limiting steps of replication restart involve the synthesis and activation of the new primer downstream. We also find little support for the concept that polymerase uncoupling, where extensive lagging-strand synthesis proceeds downstream in the absence of leading-strand synthesis, involves physical separation of the leading-strand polymerase from the replisome. Instead, our data indicate that extensive uncoupled replication likely results from a failure of the leading-strand polymerase still associated with the DNA helicase and the lagging-strand polymerase that are proceeding downstream to reinitiate synthesis.
authors.affiliation: Molecular Biology Program, Memorial Sloan-Kettering Cancer Center, 1275 York Avenue, New York, NY 10065, USA.
authors.forename: Joseph T P
authors.initials: JT
authors.lastname: Yeeles
authors.affiliation: Molecular Biology Program, Memorial Sloan-Kettering Cancer Center, 1275 York Avenue, New York, NY 10065, USA. Electronic address: kmarians@sloankettering.edu.
authors.forename: Kenneth J
authors.initials: KJ
authors.lastname: Marians
mesh_terms: D001426:Bacterial Proteins / Q000378:metabolism; D004249:DNA Damage; D004258:DNA Polymerase III / Q000378:metabolism; D019915:DNA Primase / Q000378:metabolism; D042522:DNA Replication Timing; D004269:DNA, Bacterial / Q000096:biosynthesis*; D004259:DNA-Directed DNA Polymerase / Q000235:genetics / Q000378:metabolism*; D053488:DnaB Helicases / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D007700:Kinetics; D009097:Multienzyme Complexes / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D013698:Templates, Genetic; D000099272:Replisomes
chemical_list: D001426:Bacterial Proteins; D004258:DNA Polymerase III; D019915:DNA Primase; D004269:DNA, Bacterial; D004259:DNA-Directed DNA Polymerase; D053488:DnaB Helicases; D009097:Multienzyme Complexes; D000099272:Replisomes; C103076:DnaX protein, Bacteria; C506730:dnaB protein, E coli