DNA helicase E and DNA polymerase ∈ functionally interact for displacement synthesis

John Turchi, Gregg Siegal, Robert A. Bambara

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

A functional interaction between DNA helicase E and DNA polymerase ∈ from calf thymus has been detected which results in the extension of an upstream 3′ OH through a downstream primer to the end of a synthetic template. DNA synthesis resulting in full-length extension products was dependent on the addition of DNA helicase E and hydrolysis of ATP, suggesting that displacement of the downstream primer was required. Identical reactions using DNA polymerases α and δ in place of DNA polymerase ∈ showed no full-length products dependent on helicase E, indicating that polymerases α and δ were incapable of functionally interacting with the helicase. The reaction leading to full-length extension products was time dependent and dependent on the concentration of added polymerase ∈ and helicase E. Exonucleolytic degradation of the downstream primer, or ligation of the downstream primer to the upstream 3′ OH, were not responsible for the full-length products observed. Displacement of the downstream primer by DNA helicase E was not affected by the addition of polymerase ∈ to the reactions. Template dilution experiments demonstrated that DNA polymerase ∈ and helicase E were acting in concert to perform displacement synthesis. Additional evidence for functional coordination was obtained by demonstration that DNA helicase E stimulated DNA polymerase ∈ in a standard DNA synthetic assay using dA3000·dT16 as the template-primer. The results presented are consistent with the hypothesis that DNA helicase E and DNA polymerase ∈ are capable of coordinated activities that result in displacement synthesis. A functional interaction of this sort may be involved at the eukaryotic replication fork or in DNA repair.

Original languageEnglish (US)
Pages (from-to)9008-9015
Number of pages8
JournalBiochemistry
Volume31
Issue number37
StatePublished - 1992
Externally publishedYes

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DNA Helicases
DNA-Directed DNA Polymerase
DNA
Thymus
DNA Repair
Thymus Gland
Dilution
Ligation
Hydrolysis
Assays
Repair
Demonstrations
Adenosine Triphosphate
Degradation

ASJC Scopus subject areas

  • Biochemistry

Cite this

DNA helicase E and DNA polymerase ∈ functionally interact for displacement synthesis. / Turchi, John; Siegal, Gregg; Bambara, Robert A.

In: Biochemistry, Vol. 31, No. 37, 1992, p. 9008-9015.

Research output: Contribution to journalArticle

Turchi, John ; Siegal, Gregg ; Bambara, Robert A. / DNA helicase E and DNA polymerase ∈ functionally interact for displacement synthesis. In: Biochemistry. 1992 ; Vol. 31, No. 37. pp. 9008-9015.
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