Escherichia coli replication factor Y, a component of the primosome, can act as a DNA helicase.
The primosome is a mobile multienzyme DNA replication-priming complex that requires seven Escherichia coli proteins for assembly (the products of the dnaB, dnaC, dnaG, and dnaT genes as well as proteins n and n" and replication factor Y). It has been shown previously that the primosome, in combination with the E. coli DNA polymerase III holoenzyme, can form replication forks in vitro that move at rates similar to those measured in vivo and that the primosome and one of the components of the primosome, the DNA B protein, have DNA helicase activity. Evidence is presented here that another component of the primosome, replication factor Y, possesses DNA helicase activity as well. Factor Y helicase activity requires the presence of E. coli single-stranded DNA binding protein, Mg2+, and hydrolyzable ATP or dATP. Helicase activity is stimulated 15-fold when the enzyme is actively loaded onto single-stranded DNA through a primosome assembly site, and duplex DNA is unwound unidirectionally, 3'----5', along the DNA strand to which the protein is bound.
Lee
M S
MS
Graduate Program in Molecular Biology, Memorial Sloan-Kettering Cancer Center, New York, NY 10021.
Marians
K J
KJ
D000251:Adenosine Triphosphatases / Q000502:physiology*; D001665:Binding Sites; D004265:DNA Helicases / Q000235:genetics*; D004261:DNA Replication; D004277:DNA, Single-Stranded / Q000235:genetics; D004268:DNA-Binding Proteins / Q000502:physiology*; D053488:DnaB Helicases; D009097:Multienzyme Complexes / Q000502:physiology; D012045:Regulatory Sequences, Nucleic Acid
D004277:DNA, Single-Stranded; D004268:DNA-Binding Proteins; D009097:Multienzyme Complexes; D000251:Adenosine Triphosphatases; C506730:dnaB protein, E coli; D004265:DNA Helicases; D053488:DnaB Helicases