Fluorescence (Förster) resonance energy transfer imaging of oncogene activity in living cells

A hallmark of cancer cells is their uncontrolled activation of growth signal transduction cascades comprised of oncogene products. Overexpression and activating mutations of the growth factor receptors Ras and Raf are frequently observed in human cancer cells. Several research groups, including our own, have been developing probes based on the principle of fluorescence (Förster) resonance energy transfer (FRET) to visualize how signaling molecules, including oncogene products, are regulated in normal and cancerous cells in the living state. In this review, we will briefly introduce the principle of FRET‐based probes, present an overview of the probes reported to date, and discuss the perspectives of these probes and fluorescent imaging systems in cancer biology. (Cancer Sci 2006; 97: 8–15)

[1]  Borivoj Vojnovic,et al.  Monitoring conformational changes of proteins in cells by fluorescence lifetime imaging microscopy. , 2003, The Biochemical journal.

[2]  J. Troppmair,et al.  The ins and outs of Raf kinases. , 1994, Trends in biochemical sciences.

[3]  Elliot L. Botvinick,et al.  Visualizing the mechanical activation of Src , 2005, Nature.

[4]  R. Tsien,et al.  Fluorescent indicators for Ca2+based on green fluorescent proteins and calmodulin , 1997, Nature.

[5]  A. Miyawaki,et al.  A Pair of Fluorescent Resonance Energy Transfer-based Probes for Tyrosine Phosphorylation of the CrkII Adaptor Protein in Vivo * 210 , 2001, The Journal of Biological Chemistry.

[6]  Takeharu Nagai,et al.  Shift anticipated in DNA microarray market , 2002, Nature Biotechnology.

[7]  M. Rosen,et al.  Rational design of genetically encoded fluorescence resonance energy transfer-based sensors of cellular Cdc42 signaling. , 2003, Biochemistry.

[8]  Michiyuki Matsuda,et al.  A FRET-based probe for epidermal growth factor receptor bound non-covalently to a pair of synthetic amphipathic helixes. , 2005, Experimental cell research.

[9]  Masatoshi Hagiwara,et al.  A fluorescent indicator for visualizing cAMP-induced phosphorylation in vivo , 2000, Nature Biotechnology.

[10]  Ira,et al.  Nanoscale Organization of Multiple GPI-Anchored Proteins in Living Cell Membranes , 2004, Cell.

[11]  A. Kenworthy,et al.  High-resolution FRET microscopy of cholera toxin B-subunit and GPI-anchored proteins in cell plasma membranes. , 2000, Molecular biology of the cell.

[12]  C. Coban,et al.  Interleukin-1 receptor-associated kinase-1 plays an essential role for Toll-like receptor (TLR)7- and TLR9-mediated interferon-α induction , 2005, The Journal of experimental medicine.

[13]  M. Power.  Prey exchange between a stream and its forested watershed elevates predator densities in both habitats. , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[14]  Roger Y. Tsien,et al.  Seeing the Machinery of Live Cells , 1998, Science.

[15]  M. Matsuda,et al.  RalA activation at nascent lamellipodia of epidermal growth factor-stimulated Cos7 cells and migrating Madin-Darby canine kidney cells. , 2004, Molecular biology of the cell.

[16]  K. Honda,et al.  Spatiotemporal regulation of MyD88–IRF-7 signalling for robust type-I interferon induction , 2005, Nature.

[17]  H. Kitayama,et al.  A ras-related gene with transformation suppressor activity , 1989, Cell.

[18]  R Y Tsien,et al.  Genetically encoded reporters of protein kinase A activity reveal impact of substrate tethering , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[19]  D. Hanahan,et al.  The Hallmarks of Cancer , 2000, Cell.

[20]  A. Nicholson,et al.  Mutations of the BRAF gene in human cancer , 2002, Nature.

[21]  A. Miyawaki.  Visualization of the spatial and temporal dynamics of intracellular signaling. , 2003, Developmental cell.

[22]  A. Miyawaki,et al.  Spatio-temporal images of growth-factor-induced activation of Ras and Rap1 , 2001, Nature.

[23]  Michiyuki Matsuda,et al.  Ras binding opens c‐Raf to expose the docking site for mitogen‐activated protein kinase kinase , 2005, EMBO reports.

[24]  Roger Y. Tsien,et al.  Creating new fluorescent probes for cell biology , 2003, Nature Reviews Molecular Cell Biology.

[25]  S. Nada,et al.  Mechanism of Csk-mediated Down-regulation of Src Family Tyrosine Kinases in Epidermal Growth Factor Signaling* , 2004, Journal of Biological Chemistry.

[26]  Y. Umezawa,et al.  Fluorescent Indicators for Akt/Protein Kinase B and Dynamics of Akt Activity Visualized in Living Cells* , 2003, Journal of Biological Chemistry.

[27]  R Y Tsien,et al.  Genetically encoded fluorescent reporters of protein tyrosine kinase activities in living cells , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[28]  H. Hanafusa.  Cell Transformation by RNA Tumor Viruses , 1977 .

[29]  M. Shipitsin,et al.  Activation of RalA is critical for Ras-induced tumorigenesis of human cells. , 2005, Cancer cell.

[30]  M. Matsuda,et al.  Analysis of the spatiotemporal activation of rho GTPases using Raichu probes. , 2006, Methods in enzymology.

[31]  M. Matsuda,et al.  Mechanism of the spatio‐temporal regulation of Ras and Rap1 , 2003, The EMBO journal.

[32]  M. Matsuda,et al.  Activation of Rac and Cdc42 Video Imaged by Fluorescent Resonance Energy Transfer-Based Single-Molecule Probes in the Membrane of Living Cells , 2002, Molecular and Cellular Biology.

[33]  M. Matsuda,et al.  Coactivation of Rac1 and Cdc42 at lamellipodia and membrane ruffles induced by epidermal growth factor. , 2003, Molecular biology of the cell.

[34]  J. Condeelis,et al.  Imaging Sites of N-WASP Activity in Lamellipodia and Invadopodia of Carcinoma Cells , 2004, Current Biology.

[35]  Shuichi Takayama,et al.  Lateral propagation of EGF signaling after local stimulation is dependent on receptor density. , 2002, Developmental cell.

[36]  T. Pawson,et al.  Proteomic, Functional, and Domain-Based Analysis of In Vivo 14-3-3 Binding Proteins Involved in Cytoskeletal Regulation and Cellular Organization , 2004, Current Biology.

[37]  Y. Umezawa,et al.  Fluorescent indicators for imaging protein phosphorylation in single living cells , 2002, Nature Biotechnology.

[38]  Xuejun Jiang,et al.  Grb2 regulates internalization of EGF receptors through clathrin-coated pits. , 2003, Molecular biology of the cell.

[39]  R. Tsien,et al.  Partitioning of Lipid-Modified Monomeric GFPs into Membrane Microdomains of Live Cells , 2002, Science.

[40]  C. Nobes,et al.  Rho, Rac, and Cdc42 GTPases regulate the assembly of multimolecular focal complexes associated with actin stress fibers, lamellipodia, and filopodia , 1995, Cell.

[41]  T. Hirano,et al.  STAT3 noncell-autonomously controls planar cell polarity during zebrafish convergence and extension , 2004, The Journal of cell biology.

[42]  M. Matsuda,et al.  Activity of Rho-family GTPases during cell division as visualized with FRET-based probes , 2003, The Journal of cell biology.

[43]  A. Sorkin,et al.  Interaction of EGF receptor and Grb2 in living cells visualized by fluorescence resonance energy transfer (FRET) microscopy , 2000, Current Biology.

[44]  C. Der,et al.  Signaling Interplay in Ras Superfamily Function , 2005, Current Biology.

[45]  M. Matsuda,et al.  Localized RhoA activation as a requirement for the induction of membrane ruffling. , 2005, Molecular biology of the cell.

[46]  K. Hahn,et al.  Localized Rac activation dynamics visualized in living cells. , 2000, Science.

[47]  E. Sahai,et al.  RHO–GTPases and cancer , 2002, Nature Reviews Cancer.

[48]  Roger Y. Tsien,et al.  A genetically encoded fluorescent reporter reveals oscillatory phosphorylation by protein kinase C , 2003, The Journal of cell biology.