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제121회 대한화학회 학술발표회, 총회 및 기기전시회 안내 Investigation of Nucleosome Using thdG-tC FRET System

2018년 2월 4일 22시 44분 26초
BIO.O-3 이곳을 클릭하시면 발표코드에 대한 설명을 보실 수 있습니다.
목 09시 : 40분
Life Chemistry - Oral Presentation in Chemistry of Life
저자 및
Ji Hoon Han, Soyoung Park*, Hiroshi Sugiyama*
Department of Chemistry, Graduate School of Science, Kyoto University, Japan
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승인 3건
The structural changes of a nucleosome, in which nucleosome is basic structural unit of eukaryotic chromatin, are important key to the understanding of the mechanism of genetic process. Förster Resonance Energy Transfer (FRET) is a useful tool to investigate structural dynamics of nucleosomes such as unfolding, unwrapping and repositioning. In the previous study, fluorophores such as Cy3 or Cy5 were conjugated to DNA base and/or histone protein via flexible linkers. However, rotational freedom of these dyes is a drawback for accurate analysis of the conformational dynamics. Very recently, we have developed a novel nucleic acid-based FRET system that consists of 2-aminothieno[3,4-d]pyrimidine G-mimic deoxyribonucleoside (thdG) as a donor and 1,3-diaza-2-oxophenothiazine (tC) as an acceptor. In this study, we incorporated our FRET pair into 601 sequence DNA and observed FRET efficiency of thdG and tC-containing nucleosome to investigate folding process of nucleosomes. As considering location of fluorophores in nucleosome, we designed primers containing donor and acceptor and successfully incorporated into 601 sequences DNA by PCR amplification. We found that thdG-tC FRET pair-containing 601 sequence DNA could be successfully reconstituted to nucleosomes which have a different FRET efficiencies.