【報告題目】Correlative FRET imaging for studying molecular interaction in living cells

【報 告 人】Dr. Yingxiao Wang（王英曉）

美國加利福尼亞大學聖迭戈分校（University of California, San Diego👱，UCSD）

【報告時間】2013年6月5日  下午 1：30

【報告地點】生物藥學樓3-105

【聯 系 人】力學生物學研究所 齊穎新， This e-mail address is being protected from spambots. You need JavaScript enabled to view it.

【報告人簡介】王英曉博士現任加利福尼亞大學聖迭戈分校（UCSD）生物工程系（Dept. of Bioengineering, University of California, San Diego）副教授🉐，從事力學生物學、整合生物學、新型分子傳感器、分子工程和細胞分子生物學研究，在熒光共振能量轉移（FRET） 技術發展和應用方面有卓越的貢獻🤸🏿‍♀️，在Nature🧙‍♀️、Nature Communications和PNAS等國際著名期刊發表論文60余篇，獲美國NSF青年科學家獎🦧🧜🏼‍♀️、NIH獨立科學家獎、Grainger獎和Xerox獎等。

【報告內容（摘要）】The spatiotemporal coupling between molecular activities and local functional outcomes at subcellular regions largely governs cellular physiology. However, the dynamic coordination between kinase activities and functional outcomes such as focal adhesion (FA) disassembly at the subcellular FA sites remains elusive. Here, we have developed a correlative FRET imaging method to quantify the subcellular coupling between Src kinase activation and FA disassembly at cell periphery locations in live cells. Regression analysis demonstrated that the amount of FA disassembled at cell periphery was linearly dependent on the level of Src kinase activation in fibroblasts stimulated with the platelet-derived growth factor (PDGF). Further analysis revealed that the amount of FA disassembly per unit of Src activation, and the dynamic coupling between Src activation and FA disassembly, was regulated by the concentration of fibronectin (FN) where cells were seeded. The coupling between Src activation and FA disassembly increased when integrin was inhibited, evidenced by an increased amount of FA disassembly per unit of Src activation as well as shortening of the time lag between Src activation and FA disassembly. In contrast, the inhibition of v caused a decoupling of Src activation and FA disassembly. Thus, distinct spatiotemporal coupling between Src activation and the disassembly of two FA populations mediated by different integrin subtypes was revealed by our live-cell correlation analysis approach. Our work hence highlights the power of computational bio-imaging in single live cells in deciphering the complex and noisy signaling network underlying dynamic cellular processes.