Open AccessConversion of ϕX174 Viral DNA to Double-Stranded Form by Purified Escherichia coli Proteins
Author(s) -
Sue Wickner,
Jerard Hurwitz
Publication year - 1974
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.71.10.4120
Subject(s) - dnab helicase , dnag , dna clamp , dna polymerase ii , dna replication , primase , dna polymerase , dna , dnaa , biology , microbiology and biotechnology , chemistry , biochemistry , circular bacterial chromosome , eukaryotic dna replication , gene , helicase , rna , reverse transcriptase
TheE. coli proteins that catalyze the conversion of ϕX174 single-stranded DNA to duplex DNA have now been purified extensively. The reaction depends ondnaB, dnaC(D), dnaE , anddnaG gene products, DNA elongation factors I and II,E. coli DNA binding protein, and two additionalE. coli proteins, replication factors X and Y. DNA synthesis by these proteins requires ϕX174 viral DNA, dNTPs, Mg+2 , and ATP. The product synthesized is full-length linear ϕX174 DNA. The reaction has been resolved into two steps. The first step involves the interaction of ATP and ϕX174 DNA withdnaB anddnaC(D) gene products,E. coli DNA binding protein, and replication factors X and Y in the absence of dNTPs. Subsequent dNMP incorporation requires the addition of DNA polymerase III, DNA elongation factors I and II,dnaG gene product, and dNTPs.