Quantitative analysis of transcription factor captured by intermediate filament

2022 Fiscal Year Final Research Report

• Project/Area Number: 19K22092
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Review Section: Medium-sized Section 27:Chemical engineering and related fields
• Research Institution: National Institute of Advanced Industrial Science and Technology
• Principal Investigator: Nakamura Chikashi 国立研究開発法人産業技術総合研究所, 生命工学領域, 連携研究室付 (40357661)
• Co-Investigator(Kenkyū-buntansha): 山岸 彩奈 国立研究開発法人産業技術総合研究所, 生命工学領域, 研究員 (00778293)
• Project Period (FY): 2019-06-28 – 2023-03-31
• Keywords: 中間径フィラメント / AFM / ナノニードル / 転写制御

Outline of Final Research Achievements

Intermediate filaments (IFs) have recently been reported to bind to various proteins in the cytoplasm and regulate their function. We hypothesize that IFs act as scaffolds that trap transcription-related factors and regulate gene expression by preventing nuclear translocation. In this study, we developed a method to detect IF-trapped molecules in living cells and to elucidate the mechanism of transcriptional regulation by IFs. We used antibody-modified nanoneedles and atomic force microscopy to target the transcription-related protein PHB2, which is trapped by IF vimentin, and to mechanistically detect IF-binding molecules in living cells.

Free Research Field

ナノ細胞工学

Academic Significance and Societal Importance of the Research Achievements

本研究で開発した技術により、さまざまな細胞骨格に結合するタンパク質を力学的に検出可能であると考えられ、非標識の生きた細胞に対してこれを適用することで、転写関連因子の捕縛・放出のダイナミックな細胞内の変化を解析する手法になる。これにより、細胞骨格タンパク質が支配する細胞機能調節機構を明らかにできると期待される。