Reconstitution of mitotic chromatids with a minimum set of purified proteins

2014 Fiscal Year Final Research Report

• Project/Area Number: 24770172
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Molecular biology
• Research Institution: The Institute of Physical and Chemical Research
• Principal Investigator: KEISHI Shintomi 独立行政法人理化学研究所, 平野染色体ダイナミクス研究室, 研究員 (60462694)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Keywords: 染色体 / コンデンシン / トポイソメラーゼⅡ / ヒストン / ヒストンシャペロン / 再構成 / 分裂期 / 細胞周期

Outline of Final Research Achievements

Mitotic chromatids assembly helps guarantee error-free transmission of the genome during cell division. To understand mechanistically how chromatids might be assembled, I have aimed to reconstitute them with as few purified proteins as possible in a test tube. Intensive surveys on ingredients of the assay, which are based on the hypothesis that proper targeting of major chromosomal proteins to sperm DNA is the key for the reconstitution, enable me to recapitulate the chromatid assembly process using only six protein factors (core histones, topoisomerase II, condensin I, nucleoplasmin, Nap1, FACT) and ATP. These results indicate that chromatid assembly during mitosis hinges on much simpler mechanisms than previously thought. Equally important, the chromatid reconstitution system established in this study will undoubtedly open a new avenue of dissecting specific functions of individual factors as well as their cooperation in the future.

Free Research Field

細胞生物学・生化学