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2021 Fiscal Year Final Research Report

Cryo-EM analysis using chromatin substrate film on the grid

Research Project

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Project/Area Number 19K06522
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Review Section Basic Section 43020:Structural biochemistry-related
Research InstitutionThe University of Tokyo

Principal Investigator

Takizawa Yoshimasa  東京大学, 定量生命科学研究所, 准教授 (00434291)

Project Period (FY) 2019-04-01 – 2022-03-31
Keywordsクロマチン / ヌクレオソーム / クライオ電子顕微鏡
Outline of Final Research Achievements

In eukaryotic cells, the genomic DNA is tightly packed into the nucleus by forming the chromatin. The basic repeating unit of chromatin is nucleosome, which contains ~147 base pairs of DNA and two copies of four core histones, H2A, H2B, H3, and H4.. Recently, cryo-EM analysis is an indispensable technique for structural biology. However it is still challenging to reproducibly obtain the suitable cryo-EM grid of chromatin sample for the single particle analysis. To efficiently prepare the suitable grid of chromatin sample for single-particle cryo-EM analysis, I have attempted to generate the chromatin substrate film grid, which is directly attached the chromatin on the cryo-EM grid. In this approach, I covered a single layer graphene film on the cryo-EM grid and applied the nucleosome. I successfully concentrated the nucleosome on the cryo-EM grid with a graphene film. It will be useful for further structural studies of the chromatin.

Free Research Field

構造生物学

Academic Significance and Societal Importance of the Research Achievements

本研究課題は、クロマチン基盤膜グリッドを作製することにより、クライオ電子顕微鏡によるクロマチン構造解析の効率化および高品質化を図ることを目的としている。本研究を行った結果、グラフェンシートを貼ったグリッド上で、ヌクレオソームを化学固定することなく粒子の崩壊を防ぎ、クライオ電顕で観察することが可能となった。本研究により、クロマチン基盤膜グリッドの基礎ができたことにより、ヌクレオソームおよび高次クロマチンの構造解析だけでなく、エピゲノム情報を導入したクロマチンやクロマチン結合因子複合体などの構造解析の促進が期待できる。

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Published: 2023-01-30  

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