Open AccessPriming of DNA synthesis by diadenosine 5',5"'-P1,P4-tetraphosphate with a double-stranded octadecamer as a template and DNA polymerase alpha.
Author(s) -
Paul C. Zamecnik,
Eliezer Rapaport,
Earl F. Baril
Publication year - 1982
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.79.6.1791
Subject(s) - dna polymerase , primer (cosmetics) , dna synthesis , dna , microbiology and biotechnology , biology , polymerase , dna polymerase ii , dna clamp , biochemistry , stereochemistry , chemistry , polymerase chain reaction , reverse transcriptase , gene , organic chemistry
Diadenosine 5',5"'-P1,P4-tetraphosphate (Ap4A) primes DNA synthesis in an in vitro system containing purified HeLa cell DNA polymerase alpha, deoxyadenosine triphosphate, and the double-stranded synthetic octadecamer template 5'-d-(G-G-A-G-G-C-T-T-T-T-T-T-G-G-A-G-G-C) (C-C-T-C-C-G-A-A-A-A-A-A-C-C-T-C-C-G)-d-5'; this octadecamer sequence is part of the origin region of DNA synthesis in simian virus 40. Ap4A is shown to be covalently linked to the first residue of the short deoxynucleotide chain synthesized under these experimental conditions. This template-primer system can initiate the new deoxynucleotide chain but cannot extend it beyond the A . T region.