Structural and functional analysis of chromatin assembly factor CAF-I

2019 Fiscal Year Final Research Report

• Project/Area Number: 17H06264
• Research Category: Grant-in-Aid for Challenging Research (Pioneering)
• Allocation Type: Single-year Grants
• Research Field: Biomedical structure and function and related fields
• Research Institution: High Energy Accelerator Research Organization
• Principal Investigator: Senda Toshiya 大学共同利用機関法人高エネルギー加速器研究機構, 物質構造科学研究所, 教授 (30272868)
• Project Period (FY): 2017-06-30 – 2020-03-31
• Keywords: 構造生物学 / クロマチン / 複製 / 複合体

Outline of Final Research Achievements

A large scale purification system of CAF-I complexes was established and the purified complexes were observed by cryo-EM. The results suggested that the CAF-I complexes disassembled during grid preparation. Subsequently, in order to stabilize the subunit interactions, cross-linking and gel filtration experiments were performed, but the appropriate conditions could not be found. Next, since the dissociation constants of CAF-I and histone H3/H4 are the nanomolar order, the H3/H4 is expected to stabilize CAF-I complex. Then, CAF-I-H3/H4 complexes were purified by gel filtration column. The cryo-EM experiments are in progress, but no clear particle image has been obtained yet.

Free Research Field

構造生物学

Academic Significance and Societal Importance of the Research Achievements

本研究計画が対象とするCAF-I複合体は、細胞が獲得した機能を維持する役割を果たす。ということは、CAF-I複合体の機能を阻害することができれば、細胞機能をリセットできる可能性がある。iPS細胞では、転写反応を制御することで細胞機能をリセットし、多能性幹細胞を創出しているが、CAF-I複合体の阻害は複製反応を制御することになる。本研究の目的が達成されれば、これまでとは全く異なる原理によって多能性幹細胞を創出できることが期待される。