Two-color photoactivatable probe for selective tracking of proteins and cells.
We report the development and application of photoactivatable Green Cherry (G(PA)C), the first genetically encoded "continuously red-photoactivatable green" two-color probe for live cell imaging. G(PA)C is unique in that it enables real-time tracking of selected subpopulations of proteins and organelles in the cell or of cells within tissues and whole organisms, with constant reference to the entire population of the probe. Using G(PA)C-zyxin as proof of utility, we obtained new insights into the dynamic movement of the cytoskeletal protein zyxin. We show that zyxin is continuously and rapidly recruited from the cytosol into established focal adhesions. It can also move rapidly within a given focal adhesion and "hop" between adjacent focal adhesions, emphasizing the dynamic nature of proteins within these structures. The in vivo utility of G(PA)C is exemplified by tracking hemocyte movements using a versatile transgenic Drosophila model engineered to express G(PA)C in tissues and cells of interest under the control of the GAL4-inducible promoter.
Edinburgh Cancer Research Centre, Institute of Genetics and Molecular Medicine, University of Edinburgh, Crewe Road South, Edinburgh EH4 2XR, Scotland, United Kingdom. a.welman@ed.ac.uk
Arkadiusz
A
Welman
Alan
A
Serrels
Valerie G
VG
Brunton
Mark
M
Ditzel
Margaret C
MC
Frame
D000818:Animals; D030801:Animals, Genetically Modified; D002448:Cell Adhesion; D002465:Cell Movement; D003116:Color; D029721:Drosophila Proteins / Q000737:chemistry*; D004331:Drosophila melanogaster; D022001:Focal Adhesions; D005818:Genetic Engineering; D006434:Hemocytes / Q000166:cytology; D018398:Homeodomain Proteins / Q000737:chemistry*; D008027:Light; D008164:Luminescent Proteins / Q000032:analysis*; D008853:Microscopy / Q000379:methods*; D057054:Molecular Imaging / Q000295:instrumentation* / Q000379:methods*; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D060589:Zyxin
D029721:Drosophila Proteins; D018398:Homeodomain Proteins; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; C499520:Zyx protein, Drosophila; D060589:Zyxin