Open AccessConversion of ϕX174 and fd Single-Stranded DNA to Replicative Forms in Extracts of Escherichia coli
Author(s) -
Reed B. Wickner,
Michel Wright,
Sue Wickner,
Jerard Hurwitz
Publication year - 1972
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.69.11.3233
Subject(s) - dnag , dnab helicase , primase , dna polymerase , dna polymerase ii , microbiology and biotechnology , ribonucleotide reductase , dna replication , dnaa , biology , dna clamp , dna , biochemistry , chemistry , gene , circular bacterial chromosome , eukaryotic dna replication , helicase , rna , protein subunit , reverse transcriptase
ϕX174 and M13 (fd) single-stranded circular DNAs are converted to their replicative forms by extracts ofE. coli pol A1 cells. We find that the ϕX174 DNA-dependent reaction requires Mg++ , ATP, and all four deoxynucleoside triphosphates, but not CTP, UTP, or GTP. This reaction also involves the products of thedna C,dna D,dna E (DNA polymerase III), anddna G genes, but not that ofdna F (ribonucleotide reductase). Thein vitro conversion of fd single-stranded DNA to the replicative form requires all four ribonucleoside triphosphates, Mg++ , and all four deoxynucleoside triphosphates. The reaction involves the product of genedna E but not those of genesdna C,dna D,dna F, ordna G. The reaction with fd DNA is inhibited by rifampicin or antibody to RNA polymerase, while the reaction with ϕX174 DNA is not affected by either. With the ϕX174 DNA-dependent reaction, activities have been detected that specifically complement extracts ofdna A,dna B,dna C,dna D, ordna G mutants.