
Replication of adenovirus DNA-protein complex with purified proteins.
Author(s) -
JohE Ikeda,
Takemi Enomoto,
Jerard Hurwitz
Publication year - 1981
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.78.2.884
Subject(s) - dna polymerase ii , dna polymerase , microbiology and biotechnology , biology , dna replication , dna , dna clamp , replication factor c , eukaryotic dna replication , aphidicolin , replication protein a , dna polymerase i , adenoviridae , control of chromosome duplication , dna binding protein , biochemistry , recombinant dna , rna , gene , reverse transcriptase , transcription factor
A protein fraction isolated from the cytosol of adenovirus-infected HeLa cells, which contained DNA polymerase alpha, catalyzed adenoviral DNA replication in the presence of adenovirus DNA binding protein, eukaryotic DNA polymerase beta, ATP, all four dNTPs, and MgCl2. DNA replication started at either end of exogenously added adenoviral DNA and was totally dependent on the presence of terminal 55,000-dalton proteins on the DNA template. The replicaton of adenovirus DNA in the system was sensitive to aphidicolin and retained nearly all the properties of DNA replication that occur in vivo or in vitro with crude extracts. The 5'-ends of the newly synthesized adenovirus DNA strands were covalently linked to an 80,000-dalton protein linked to dCMP. DNA synthesized with purified proteins was only 25-50+ the length of parental viral strands. Addition of cytosol extracts from uninfected HeLa cells to reaction mixtures containing purified proteins gave full-length adenoviral DNA strands.