Small interfering RNAs as a tool to assign Rho GTPase exchange-factor function in vivo.
Rho GTPases control a complex network of intracellular signalling pathways. Whereas progress has been made in identifying downstream signalling partners for these proteins, the characterization of Rho upstream regulatory guanine-nucleotide exchange factors (GEFs) has been hampered by a lack of suitable research tools. Here we use small interfering RNAs (siRNAs) to examine the cellular regulation of the RhoB GTPase, and show that RhoB is activated downstream of the epidermal-growth-factor receptor through the Vav2 exchange factor. These studies demonstrate that siRNAs are an ideal research tool for the assignment of Rho GEF function in vivo.
Mammalian Cell Biology Laboratory, Department of Biochemistry, School of Medical Sciences, University of Bristol, Bristol BS8 1TD, UK.
Alexandra
A
Gampel
Harry
H
Mellor
D003001:Cloning, Molecular; D004789:Enzyme Activation; D004815:Epidermal Growth Factor / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D020662:Guanine Nucleotide Exchange Factors / Q000378:metabolism; D007700:Kinetics; D015513:Oncogene Proteins / Q000378:metabolism; D034741:RNA, Small Interfering; D022661:RNA, Untranslated / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D020741:rho GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism*
D020662:Guanine Nucleotide Exchange Factors; D015513:Oncogene Proteins; D034741:RNA, Small Interfering; D022661:RNA, Untranslated; D011993:Recombinant Fusion Proteins; D004815:Epidermal Growth Factor; D020741:rho GTP-Binding Proteins