PMID:8663105

Citation	Ng, JY and Marians, KJ (1996) The ordered assembly of the phiX174-type primosome. II. Preservation of primosome composition from assembly through replication. J. Biol. Chem. 271:15649-55
Abstract	Gel filtration chromatography was used to isolate both preprimosomal and primosomal complexes formed on single-stranded DNA-binding protein-coated phiX174 DNA by the combination of PriA, PriB, PriC, DnaT, DnaB, DnaC, and DnaG. The presence and relative amounts of primosomal proteins in these complexes were determined by Western blotting. Protein-DNA complexes isolated (i) after assembly in the presence of 10 microM ATP, (ii) after preprimosome movement in the presence of 1 mM ATP, (iii) after priming in the presence of the four ribonucleoside triphosphates, or (iv) after complementary strand DNA replication in the presence of the DNA polymerase III holoenzyme all had the same protein composition; preprimosomes contained PriA, PriB, PriC, DnaT, and DnaB, whereas primosomes included DnaG. The stable association of DnaG with the protein-DNA complex could be attributed partially to its ability to remain bound to the primers synthesized. In the absence of PriC, the efficiencies of priming and replication were reduced by one-third and one-half, respectively, even though PriC was not required for the formation of stable protein-DNA complexes on a 304-nucleotide-long single strand of DNA containing a primosome assembly site (Ng, J. Y., and Marians, K. J. (1996) J. Biol. Chem. 271, 15642-15648). We hypothesize that maintenance of the primosome on the replicated DNA may provide a mechanism to allow primosomes to participate in the resolution of recombination intermediates and intermediates formed during double strand break repair by permitting the re-establishment of a replication fork.
Links	PubMed
Keywords	Bacterial Proteins/metabolism; Bacteriophage phi X 174/genetics; Bacteriophage phi X 174/ultrastructure; DNA/metabolism; DNA Primase; DNA Replication; DNA, Single-Stranded/metabolism; DNA, Viral/metabolism; DNA-Binding Proteins/metabolism; Deoxyribonucleoproteins/metabolism; Escherichia coli; Macromolecular Substances; RNA Nucleotidyltransferases/metabolism; RNA, Viral/metabolism; Replication Protein A
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
ECOLI:DNAG	enables	GO:0005515: protein binding	ECO:0000353: physical interaction evidence used in manual assertion	UniProtKB:P0ACB0	F	occurs_in:(GO:0043234)	Seeded From UniProt	complete
ECOLI:DNAB	enables	GO:0005515: protein binding	ECO:0000353: physical interaction evidence used in manual assertion	UniProtKB:P0ABS5	F	occurs_in:(GO:0043234)	Seeded From UniProt	complete
ECOLI:DNAB	enables	GO:0005515: protein binding	ECO:0000353: physical interaction evidence used in manual assertion	UniProtKB:P0AEF0	F	occurs_in:(GO:0043234)	Seeded From UniProt	complete
ECOLI:DNAB	enables	GO:0005515: protein binding	ECO:0000353: physical interaction evidence used in manual assertion	UniProtKB:P23862	F		Seeded From UniProt	complete
ECOLI:DNAC	enables	GO:0005515: protein binding	ECO:0000353: physical interaction evidence used in manual assertion	UniProtKB:P0ACB0	F	occurs_in:(GO:0043234)	Seeded From UniProt	complete
ECOLI:PRIC	enables	GO:0005515: protein binding	ECO:0000353: physical interaction evidence used in manual assertion	UniProtKB:P0ACB0	F		Seeded From UniProt	complete
ECOLI:PRIC	involved_in	GO:0006270: DNA replication initiation	ECO:0000314: direct assay evidence used in manual assertion		P		Seeded From UniProt	complete

References

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