engleski

Illumination of coronin function by the expression of green fluorescent protein (GFP)-tagged coronin in Dictyostelium discoideum

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. The phosphorylation of actin at tyrosine 53 is eight times higher in coronin null mutants than in wild type. 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 has shown that coronin is found in variety of dynamic F-actin rich structures derived from the cell cortex. In chemotactic cells coronin is recruited to the leading edge. GFP-tagged coronin complements actin phosphorylation at tyrosine 53 in coronin null cells. The other defects that characterize coronin null phenotype appear to be partially complemented by GFP-tagged coronin.

coronin; cytockeleton; GFP

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