| Abstract: |
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 promosome 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 raection 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. |
| Keywords: |
dna binding protein; nonhuman; dna replication; bacterial proteins; escherichia coli; base sequence; helicase; single stranded dna; multienzyme complexes; dna, bacterial; dna helicases; primosome; nucleotidyltransferases; dna, circular; nucleotides, cyclic; priority journal; support, non-u.s. gov't; support, u.s. gov't, p.h.s.; adenosinetriphosphatase; mitochondrial adp, atp translocases
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