Versatile fluorescent probes for actin filaments based on the actin-binding domain of utrophin.
Actin filaments (F-actin) are protein polymers that undergo rapid assembly and disassembly and control an enormous variety of cellular processes ranging from force production to regulation of signal transduction. Consequently, imaging of F-actin has become an increasingly important goal for biologists seeking to understand how cells and tissues function. However, most of the available means for imaging F-actin in living cells suffer from one or more biological or experimental shortcomings. Here we describe fluorescent F-actin probes based on the calponin homology domain of utrophin (Utr-CH), which binds F-actin without stabilizing it in vitro. We show that these probes faithfully report the distribution of F-actin in living and fixed cells, distinguish between stable and dynamic F-actin, and have no obvious effects on processes that depend critically on the balance of actin assembly and disassembly.
Burkel
Brian M
BM
Department of Zoology, University of Wisconsin-Madison, Madison, Wisconsin, USA.
von Dassow
George
G
Bement
William M
WM
D008841:Actin Cytoskeleton / Q000378:metabolism*; D000818:Animals; D005260:Female; D005455:Fluorescent Antibody Technique / Q000379:methods; D005456:Fluorescent Dyes / Q000378:metabolism*; D006801:Humans; D008840:Microfilament Proteins / Q000378:metabolism*; D038761:Photobleaching; D017434:Protein Structure, Tertiary; D049411:Utrophin / Q000378:metabolism*; D014981:Xenopus
D005456:Fluorescent Dyes; D008840:Microfilament Proteins; D049411:Utrophin