PCNA-dependent DNA polymerase δ from rabbit bone marrow

Chang De Lu, John Byrnes

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Proliferating cell nuclear antigen (PCNA) and PCNA-dependent DNA polymerase δ were partially purified and characterized from rabbit bone marrow. Rabbit DNA polymerase δ sediments at 8 2 S upon glycerol density gradient centrifugation. Similar to calf thymus PCNA-dependent DNA polymerase δ, a 125-123-kDa doublet and 48-kDa polypeptides correlate with DNA polymerase activity. Western blotting of rabbit DNA polymerase δ with polyclonal antibody to calf thymus PCNA-dependent DNA polymerase δ gives the same results as calf thymus δ; the 125-123-kDa doublet is recognized. PCNA-dependent DNA polymerase δ is resistant to inhibition by dideoxynucleotides and is relatively insensitive to inhibition by N2[p-(n-butyl)phenyl]dGTP. A 3′→5′ exonuclease copurifies with the DNA polymerase. The processivity of DNA polymerase δ alone is very low but greatly increases with the addition of PCNA from rabbit bone marrow or calf thymus. Comparative studies of the original DNA polymerase δ from rabbit bone marrow demonstrate a lack of recognition by antibodies to calf thymus δ and a high degree of processivity in the absence of PCNA. Additionally, the originally described DNA polymerase δ is a single polypeptide of 122 kDa. These features would recategorize the original δ to the ε category by recently proposed convention. PCNA-dependent DNA polymerase δ is a relatively minor component of rabbit bone marrow compared to DNA polymerase α and PCNA-independent DNA polymerase δ (ε), the relative proportions being α, 60%; δ, 7%; and ε, 30%.

Original languageEnglish
Pages (from-to)12403-12409
Number of pages7
JournalBiochemistry
Volume31
Issue number49
StatePublished - Dec 1 1992

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Proliferating Cell Nuclear Antigen
DNA-Directed DNA Polymerase
Bone
Bone Marrow
Rabbits
Thymus
Thymus Gland
Dideoxynucleotides
Exonucleases
Peptides
Density Gradient Centrifugation
Centrifugation
Antibodies
Glycerol
Sediments

ASJC Scopus subject areas

  • Biochemistry

Cite this

PCNA-dependent DNA polymerase δ from rabbit bone marrow. / Lu, Chang De; Byrnes, John.

In: Biochemistry, Vol. 31, No. 49, 01.12.1992, p. 12403-12409.

Research output: Contribution to journalArticle

Lu, Chang De ; Byrnes, John. / PCNA-dependent DNA polymerase δ from rabbit bone marrow. In: Biochemistry. 1992 ; Vol. 31, No. 49. pp. 12403-12409.
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