Detection of histone modification cross-talk using fluorescence cross-correlation spectroscopy

2014 Fiscal Year Final Research Report

• Project/Area Number: 24657003
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Genetics/Genome dynamics
• Research Institution: Tokyo Institute of Technology (2014); Osaka University (2012-2013)
• Principal Investigator: HIROSHI Kimura 東京工業大学, 生命理工学研究科, 教授 (30241392)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Keywords: クロマチン / 染色体 / 蛍光相互相関分光法 / エピジェネティクス / 蛋白質翻訳後修飾

Outline of Final Research Achievements

Post-translational histone modification plays an important role in the regulation and maintenance of genome function, such as transcription and DNA repair. It remains unknown, however, how a combination of different modifications is regulated and functions specifically. This study aimed to develop a rapid and sensitive system to detect a combination of modifications using fluorescence cross-correlation spectroscopy. When Alexa488-labeled and Cy5-labeled antibodies are mixed with purified histone complexes, they tended to form aggregations, because the solubilization conditions of labeled antibodies and histones were different.This problem was partly solved by using antigen-binding fragments and nucleosomes, instead of IgG and histones. Additional experiments revealed that a combination of different modifications could be detected by measuring fluorescence resonance enery transfer.

Free Research Field

エピジェネティクス