Immunodetection of Rho-like plant proteins with Rac1 and Cdc42Hs antibodies |
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Authors: | Couchy I; Minic Z; Laporte J; Brown S; Satiat-Jeunemaitre B |
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Institution: | Institut des Sciences Vegetales, Laboratoire de Cytometrie, CNRS UPR40, 91198 Gif-sur-Yvette, France; Corresponding author |
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Abstract: | A few small GTP-binding proteins of the Ras superfamily have been
identified in plants, including members of the Rho family: the proteins
belonging to this group are known in mammalian and yeast cells to be
involved in the control of polarity, cell morphogenesis and movement by
regulating cytoskeleton organization. An investigation into where some
Rho-like proteins are located in plant cells was made. The antibodies used,
anti-Cdc42Hs and anti-Rac 1, were raised against conserved characteristic
sequences of Cdc42Hs and Rac1 mammalian proteins respectively. In fixed
cells, Cdc42Hs antibody recognized epitopes generally co-localized with
microtubules which may be implicated in the establishment of cell polarity,
whereas the proteins recognized by Rac1 antibody seemed to be associated
with organelle membranes. The same anti-bodies were used in Western blots
of proteins from tobacco BY-2 and lucerne A2 suspension cells: Cdc42Hs
antibody recognized three bands whereas Rac1 antibody revealed only one
band of 18 kDa Mr. A 35S]GTP overlay revealed
four bands of the same Mr as those recognized in
Western blots by Cdc42Hs and Rac1 antibodies.Key words: Rho G-proteins,
Cdc42, Rac1, Immunofluorescence, plant cells.
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