The Escherichia coli primosome can translocate actively in either direction along a DNA strand.
The primosome is a mobile multiprotein DNA replication-priming apparatus that requires seven Escherichia coli proteins (replication factor Y (protein n'), proteins n and n", and the products of the dnaB, dnaC, dnaT, and dnaG genes) for assembly at a specific site (termed a primosome assembly site) on single-stranded DNA binding protein-coated single-stranded DNA. Two of the protein components of the primosome have intrinsic DNA helicase activity. The DNA B protein acts in the 5'----3' direction, whereas factor Y acts in the 3'----5' direction. The primosome complex has DNA helicase activity when present at a replication fork in conjunction with the DNA polymerase III holoenzyme. In this report, evidence is presented that the multiprotein primosome per se can act as a DNA helicase in the absence of the DNA polymerase III holoenzyme. The primosome DNA helicase activity can be manifested in either direction along the DNA strand. The directionality of the primosome DNA helicase activity is modulated by the concentration and type of nucleoside triphosphate present in the reaction mixture. This DNA helicase activity requires all the preprimosomal proteins (the primosomal proteins minus the dnaG-encoded primase). Preprimosome complexes must assemble at a primosome assembly site in order to be loaded onto the single-stranded DNA and act subsequently as a DNA helicase. The 5'----3' primosome DNA helicase activity requires a 3' single-stranded tail on the fragment to be displaced, while the 3'----5' activity does not require a 5' single-stranded tail on the fragment to be displaced. Multienzyme preprimosomes moving in either direction are capable of associating with the primase to form complete primosomes that can synthesize RNA primers.
Lee
M S
MS
Program in Molecular Biology, Sloan-Kettering Institute, New York 10021.
Marians
K J
KJ
D000251:Adenosine Triphosphatases / Q000502:physiology; D001426:Bacterial Proteins / Q000235:genetics*; D001483:Base Sequence; D004265:DNA Helicases / Q000378:metabolism; D004269:DNA, Bacterial / Q000096:biosynthesis* / Q000502:physiology; D004270:DNA, Circular / Q000378:metabolism* / Q000502:physiology; D053488:DnaB Helicases; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics* / Q000502:physiology; D029968:Escherichia coli Proteins; D000226:Mitochondrial ADP, ATP Translocases / Q000235:genetics* / Q000502:physiology; D009097:Multienzyme Complexes / Q000502:physiology; D009712:Nucleotides, Cyclic / Q000502:physiology; D009713:Nucleotidyltransferases / Q000235:genetics*
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D004270:DNA, Circular; D029968:Escherichia coli Proteins; D009097:Multienzyme Complexes; D009712:Nucleotides, Cyclic; D000226:Mitochondrial ADP, ATP Translocases; D009713:Nucleotidyltransferases; D000251:Adenosine Triphosphatases; C027093:TraI protein, E coli; C506730:dnaB protein, E coli; D004265:DNA Helicases; D053488:DnaB Helicases