Elucidation of DNA repair pathway involving the novel endonuclease derived from hyperthermophilic archaea
| Project/Area Number |
18K05442
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Multi-year Fund |
|---|
| Section | 一般 |
|---|
| Review Section |
Basic Section 38030:Applied biochemistry-related
|
|---|
| Research Institution | Kyushu University |
|---|
Principal Investigator |
Ishino Sonoko 九州大学, 農学研究院, 准教授 (80399740)
|
|---|
| Project Period (FY) |
2018-04-01 – 2021-03-31
|
| Project Status |
Completed (Fiscal Year 2020)
|
| Budget Amount *help |
¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000)
Fiscal Year 2020: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2019: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2018: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
|
|---|
| Keywords | DNA修復 / アーキア / 遺伝子破壊 / DNA損傷 / 超好熱性アーキア / 遺伝子操作 / 遺伝子工学技術 |
|---|
| Outline of Final Research Achievements |
Base deamination is a typical form of DNA damage, and it must be repaired quickly to maintain the genome integrity of living organisms. Endonuclease V (EndoV) and Endonuclease Q (EndoQ) from hyperthermophilic archaea hydrolyze the phosphodiester bond at the 3′and 5′ site from the damaged nucleotide, respectively.
The two DNA polymerases, PolB and PolD, could repair the lesion and elongate the DNA strand. The in vitro reconstitution system was constructed using the recombinant proteins derived from hyperthermophilic archaea.
EndoMS recognizes and cleaves mismatched base pairs. In order to investigate the intracellular function of the EndoMS gene, mutation accumulation assay was performed using the gene-disrupted strains. Mutation spectra were consistent with the biochemical properties.
|
| Academic Significance and Societal Importance of the Research Achievements |
本研究は、生物の遺伝情報を担うDNAが化学的損傷を受けたとき、どのように修復し遺伝情報を守っているのかを理解することを目的に、遺伝学的、および分子化学的にDNA修復機構の解明を進めた。厳しい生育環境で自らの遺伝情報を安定に維持している超好熱性アーキアにおいて、主導するタンパク質因子がどのように関連分子を認識してDNAを修復するのか理解が深まった。
また細胞内に蓄積する変異をゲノムDNAの塩基配列を直接分析することにより、生化学的性質と合わせて議論することを可能にした。
これらの成果は基礎科学への貢献と共に、新たな遺伝子工学技術の開発および医療分野への応用にもつながる。
|
Report
(3 results)
Research Products
(34 results)
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
[Presentation] Reconstitution of endonuclease Q-mediated and endonuclease V-mediated repair pathways for deaminated base repair in the hyperthermophilic archaeon, Thermococcus kodakarensis.2019
Author(s)
Meika Harada, Mariko Nagata, Keisuke Oki, Rie Matsumi, Tamotsu Kanai, Haruyuki Atomi,Tomoyuki Numata, Miyako Shiraishi, Sonoko Ishino, and Yoshizumi Ishino
Organizer
15th International Congress on Thermophiles
Related Report
Int'l Joint Research
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
