Differential Ras Activation between Caveolae/Raft and Non-Raft Microdomains

Cell Structure and Function

Atsushi Miyawaki¹  Takashi Fukano¹  Michiyuki Matsuda²  Asako Sawano¹  Yusuke Ohba² 

[1] Laboratory for Cell Function and Dynamics, Advanced Technology Development Center, Brain Science Institute, RIKEN;Department of Tumor Virology, Research Institute for Microbial Diseases, Osaka University

关键词: Ras;    FRET;    fringe projection;   

DOI  :  10.1247/csf.06019

学科分类：分子生物学，细胞生物学和基因

来源: Japan Society for Cell Biology

【 摘 要 】

References(20)Cited-By(11)Although the consequences of Ras activation have been studied extensively in the context of oncogenesis, its regulation in physiological modes of signal transduction is not well understood. A fluorescent indicator, Raichu-Ras, was fused to the C-terminal hypervariable regions of H-Ras and K-Ras to create indicators for Ras activation within caveolae/rafts (Raichu-tH) and non-raft domains (Raichu-tK) of the plasma membrane, respectively. Raichu-tH was also found abundantly in endomembranes. To monitor Ras activation with high spatial resolution, it is imperative to observe sectioned images of the signals. We have developed a wide-field fluorescence microscope equipped with a digital micromirror device (DMD) to acquire optically sectioned images using fringe projection. This system provides reliable signals from fluorescence resonance energy transfer (FRET) between cyan and yellow mutants of green fluorescent protein. We have used this system to demonstrate that, upon stimulation with growth factors, the two indicators are activated in spatially and temporally unique patterns.

【 授权许可】

Unknown