Development of DNA damaging agents using poly(ADP-ribose) hydrolysates

2018 Fiscal Year Final Research Report

• Project/Area Number: 16K00562
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Risk sciences of radiation and chemicals
• Research Institution: Kanagawa Institute of Technology
• Principal Investigator: Takamura Takeji 神奈川工科大学, 工学部, 教授 (50342910)
• Research Collaborator: Masutani Mitsuko
• Project Period (FY): 2016-04-01 – 2019-03-31
• Keywords: 変異原の検出 / ポリADPリボース

Outline of Final Research Achievements

Poly(ADP-ribose) (PAR) is a well known polymer molecule that is produced when DNA strand scissions are formed by the action of alkylating reagents. Because PAR is easily hydrolyzed by the enzymes like phosphatase and phosphodiesterase to form ribosyl-adenosine (R-Ado). Because, quantification of R-Ado is, therefore, estimated to know the amount DNA scissions in cells, high sensitive detection system of R-Ado was required. PAR was initially purified from the cell treated with mutagens, and hydrolyzed by several enzymes to form R-Ado efficiently. Obtaiened R-ado solution was quantified by the LC/MS/MS. Amounts of R-Ado was well correlated with the activity of the mutagen that formed micronucleus in cells. Moreover, we found that formation rate of PAR is differed among several mutagens.

Free Research Field

環境発がん

Academic Significance and Societal Importance of the Research Achievements

身の回りには多くの変異・がん原性物質が存在している。バクテリアを用いる変異原性試験は、これまでにも多くの分野で用いられてきており、すでに多くのデータが蓄積されている。一方で培養細胞を用いる遺伝毒性検出系ではヒトリスクを考える上で重要であるが，結果の判定に多くの時間を費やすために労力のいる仕事である。そこで，DNAに損傷が生じたときに起こる細胞内イベントを定量することでより簡潔に，そして精度の良い方法論を構築するために，DNA損傷時に生じるポリADPリボース（PAR)の生成に着目した。生成したPARは加水分解後、小分子であるリボシルアデノシンへと変化するため、高感度の分析が可能となる。