A new DNA polymerase species from Drosophila melanogaster

A probable mus308 gene product

Masahiko Oshige, Norikazu Aoyagi, Paul V. Harris, Kenneth C. Burtis, Kengo Sakaguchi

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

Harris et al. [P.V. Harris, O.M. Mazina, E.A. Leonhardt, R.B. Case, J.B. Boyd, K.C. Burtis, Molecular cloning of Drosophila mus308, a gene involved in DNA cross-link repair with homology to prokaryotic DNA polymerase I genes, Mol. Cell. Biol., 16 (1996) 5764-5771.] reported the molecular cloning of Drosophila mus308 gene, and its nucleotide and protein sequences similar to DNA polymerase I. In the present study, we attempted to find and isolate the gene product by purifying a DNA polymerase fraction not present in mus308 flies. A new DNA polymerase with properties different from those of any known polymerase species was identified and partially purified from the wild-type fly embryos through ten column chromatographies. The enzyme was resistant to aphidicolin, but sensitive to ddTTP and NEM. Human proliferating cell nuclear antigen (PCNA) and Drosophila replication protein A (RP-A) did not affect the polymerase activity. It preferred poly(dA)/oligo(dT) as a template-primer. The molecular mass was about 230 kDa with a broad peak region of 200 to 300 kDa in HiPrep16/30 Sephacryl S-300 gel filtration. These properties a different from those of all reported Drosophila polymerase classes such as α, β, γ, δ, ε and ζ and closely resemble those of the gene product expected from the nucleotide sequence. The new polymerase species appears to have ATPase and 3'-5' exonuclease activities as shown by the chromatographies. Copyright (C) 1999 Elsevier Science B.V.

Original languageEnglish (US)
Pages (from-to)183-192
Number of pages10
JournalMutation Research - DNA Repair
Volume433
Issue number3
DOIs
StatePublished - Apr 9 1999

Fingerprint

DNA-Directed DNA Polymerase
Drosophila melanogaster
Genes
Drosophila
DNA Polymerase I
Cloning
Molecular Cloning
Diptera
Chromatography
Nucleotides
Replication Protein A
Aphidicolin
Exonucleases
Column chromatography
Proliferating Cell Nuclear Antigen
Molecular mass
Gel Chromatography
Adenosine Triphosphatases
Repair
Embryonic Structures

Keywords

  • Drosophila melanogaster
  • mus308 Mutant
  • Pol I-like DNA polymerase

ASJC Scopus subject areas

  • Toxicology
  • Genetics
  • Molecular Biology

Cite this

A new DNA polymerase species from Drosophila melanogaster : A probable mus308 gene product. / Oshige, Masahiko; Aoyagi, Norikazu; Harris, Paul V.; Burtis, Kenneth C.; Sakaguchi, Kengo.

In: Mutation Research - DNA Repair, Vol. 433, No. 3, 09.04.1999, p. 183-192.

Research output: Contribution to journalArticle

Oshige, Masahiko ; Aoyagi, Norikazu ; Harris, Paul V. ; Burtis, Kenneth C. ; Sakaguchi, Kengo. / A new DNA polymerase species from Drosophila melanogaster : A probable mus308 gene product. In: Mutation Research - DNA Repair. 1999 ; Vol. 433, No. 3. pp. 183-192.
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