Elucidating functions of DP1 and DP2 subunits from the Thermococcus kodakarensis family D DNA polymerase

Natsuki Takashima, Sonoko Ishino, Keisuke Oki, Mika Takafuji, Takeshi Yamagami, Ryotaro Matsuo, Kouta Mayanagi, Yoshizumi Ishino

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

DNA polymerase D (PolD), originally discovered in Pyrococcus furiosus, has no sequence homology with any other DNA polymerase family. Genes encoding PolD are found in most of archaea, except for those archaea in the Crenarchaeota phylum. PolD is composed of two proteins: DP1 and DP2. To date, the 3D structure of the PolD heteromeric complex is yet to be determined. In this study, we established a method that prepared highly purified PolD from Thermococcus kodakarensis, and purified DP1 and DP2 proteins formed a stable complex in solution. An intrinsically disordered region was identified in the N-terminal region of DP1, but the static light scattering analysis provided a reasonable molecular weight of DP1. In addition, PolD forms as a complex of DP1 and DP2 in a 1:1 ratio. Electron microscope single particle analysis supported this composition of PolD. Both proteins play an important role in DNA synthesis activity and in 3′–5′ degradation activity. DP1 has extremely low affinity for DNA, while DP2 is mainly responsible for DNA binding. Our work will provide insight and the means to further understand PolD structure and the molecular mechanism of this archaea-specific DNA polymerase.

Original languageEnglish
Pages (from-to)161-172
Number of pages12
JournalExtremophiles
Volume23
Issue number1
DOIs
Publication statusPublished - Jan 22 2019

Fingerprint

Thermococcus
Transcription Factor DP1
Archaea
DNA-Directed DNA Polymerase
DNA
Crenarchaeota
Pyrococcus furiosus
Sequence Homology
Molecular Structure
Molecular Weight
Electrons
Light
Genes
Proteins

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Molecular Medicine

Cite this

Elucidating functions of DP1 and DP2 subunits from the Thermococcus kodakarensis family D DNA polymerase. / Takashima, Natsuki; Ishino, Sonoko; Oki, Keisuke; Takafuji, Mika; Yamagami, Takeshi; Matsuo, Ryotaro; Mayanagi, Kouta; Ishino, Yoshizumi.

In: Extremophiles, Vol. 23, No. 1, 22.01.2019, p. 161-172.

Research output: Contribution to journalArticle

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AU - Yamagami, Takeshi

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