Rho localization in cells and tissues
Yonemura S, Hirao-Minakuchi K and Nishimura Y
Exp Cell Res 295(2):300-14 (2004)
SUMMARY
Rho family small GTPases regulate cytoskeletal organization. Although their
spatiotemporal activities appear to be important for cellular morphogenesis,
there has been little characterization of the localization of Rho family
GTPases in cells and tissues. Here we show precise localization of Rho
subfamily proteins in mammalian cultured cells and tissues through evaluation
of anti-Rho antibodies and fixation protocols. Although Rho is not a structural
protein but functions as a switching molecule, it often localizes at several
distinct domains or structures of cells. In cultured epithelial cells,
Rho was highly accumulated at lateral membranes. However, in fibroblastic
cells, Rho appeared to be distributed evenly in the cytoplasm. Rho concentration
at the cleavage furrow at cytokinesis was generally observed. In A431
cells, Rho translocation from the cytoplasm to elongating microvilli at
the apical membrane within 30 s after EGF stimulation was clearly demonstrated.
Also, Myc- or GFP-tagged RhoA did not always reflect the localization
of endogenous Rho, indicating a drawback of protein-tagging methods for
localization research. In mouse tissues, Rho localization differed depending
on cell type, probably reflecting the functional differences of each cell
type.
LINK
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