Elucidation of mechanisms of transcription cycle using multi-color, single-molecule, quantitative fluorescence imaging of living cell nuclei.

2016 Fiscal Year Final Research Report

• Project Area: Integral understanding of the mechanism of transcription cycle through quantitative, high-resolution approaches
• Project/Area Number: 24118006
• Research Category: Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Tokyo Institute of Technology
• Principal Investigator: SAKATA-SOGAWA Kumiko 東京工業大学, 生命理工学院, 准教授 (20291073)
• Co-Investigator(Renkei-kenkyūsha): TOKUNAGA Makio 東京工業大学, 生命理工学院, 教授 (00192659) ; KIMURA Hiroshi 東京工業大学, 科学技術創成研究院, 教授 (30241392) ; SAITOH Noriko 熊本大学, 発生医学研究所, 准教授 (40398235)
• Project Period (FY): 2012-06-28 – 2017-03-31
• Keywords: １分子イメージング / 超解像イメージング / 転写調節 / 遺伝子発現 / 生体分子

Outline of Final Research Achievements

Transcription cycle consists of various protein factors, which interact in different ways depending on their stage in the cycle. Single-molecule approaches provide quantitative and high-resolution analysis, through capturing heterogeneous behavior of the protein factors.
By installing a single-molecule fluorescence microscope based on our original HILO illumination method, we analyzed the interaction of transcription factors and molecular dynamics of chromatin remodeling complex. Super-resolution imaging with HILO microscopy realized high-resolution analysis in the spatial distribution of transcription-related proteins. We further developed an image-analyzing method using single-molecule tracking approach. Combining this technique with three-color simultaneous single-molecule imaging, we quantified the dynamics and kinetics of molecules. This approach facilitates the spatiotemporal quantification of the dynamics and kinetics of interactions of protein factors in transcription cycle.

Free Research Field

生物物理学