A novel DNA polymerase in the hyperthermophilic archaeon, Pyrococcus furiosus: Gene cloning, expression, and characterization

Takashi Uemori, Yoshimi Sato, Ikunoshin Kato, Hirofumi Doi, Yoshizumi Ishino

研究成果: Contribution to journalArticle

108 引用 (Scopus)

抜粋

Background: In many respects Archaea are much more like eukaryotes than prokaryotes with respect to the conservation of many of the components involved in transcription, translation and DNA replication. So far, only a few DNA polymerases with structures similar to those of eukaryotic DNA polymerase α have been found in Archaea. The identification and characterization of all the DNA polymerases of one archaeon would add considerably to our knowledge of the basic mechanisms of DNA replication in these organisms. Results: We have identified a novel DNA polymerase composed of two proteins, DP1 and DP2, with molecular weights of 69 294 Da and 143 161 Da, respectively, in the hyperthermophilic archaeon, Pyrococcus furiosus, and have cloned the corresponding genes which are tandemly arranged on the Pyrococcus genome. No significant sequence homology was found between these two proteins and other known DNA polymerases. The pol genes were transcribed as part of a single operon that additionally contained genes homologous to the cdc18+/CDC6 and Dmc1/Rad51 family of proteins. We purified the Pyrococcus DNA polymerase from Escherichia coli strains expressing the cloned genes and characterized its activity. It possesses strong 3′ → 5′ exonucleolytic activity and has a template-primer preference which is characteristic of a replicative DNA polymerase. Conclusion: In P. furiosus, we identified a second DNA polymerase encoded by two genes, neither of which display significant homology to any other known DNA polymerase. Both the enzymatic properties of the enzyme and the gene organization raise the possibility that this enzyme might be the replicative DNA polymerase of P. furiosus.

元の言語英語
ページ(範囲)499-512
ページ数14
ジャーナルGenes to Cells
2
発行部数8
DOI
出版物ステータス出版済み - 1 1 1997
外部発表Yes

All Science Journal Classification (ASJC) codes

  • Genetics
  • Cell Biology

フィンガープリント A novel DNA polymerase in the hyperthermophilic archaeon, Pyrococcus furiosus: Gene cloning, expression, and characterization' の研究トピックを掘り下げます。これらはともに一意のフィンガープリントを構成します。

  • これを引用