engleski

Illumination of actin cytoskeleton using green fluorescent protein

Coronin is an actin-associated protein originally isolated from Dictyostelium discoideum. It belongs to the WD-repeat family of proteins. Dictyostelium mutants lacking coronin have marked defects in cell locomotion, cytokinesis, phagocytosis and pinocytosis. 5'-end of GFP gene was fused at 3'-end of coronin gene and vice versa. The fused genes were cloned into pDBsrH vector that contains blasticidin S resistance cassette. D. discoideum wild type cells and coronin null cells were transformed with recombinant vector by electroporation. The transformants that expressed GFP-tagged coronin were identified by fluorescent microscopy. Direct localization using GFP-tagged coronin has shown that coronin is found in a variety of dynamic F-actin rich structures derived from the cell cortex. In chemotactic cells coronin is recruited to the leading edge. The defects that characterize coronin null phenotype appear to be partially complemented by GFP-tagged coronin.

coronin; actin cytoskeleton; Dictyostelium discoideum; GFP

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