Different functions of Smg GDP dissociation stimulator and mammalian counterpart of yeast Cdc25.
We have previously shown that both Smg GDP dissociation stimulator (GDS) and mammalian Cdc25 (mCdc25) stimulate the GDP/GTP exchange reaction of Ki-Ras and that Smg GDS is active only on the post-translationally lipid-modified form of Ki-Ras, whereas mCdc25 is active on both the lipid-modified and unmodified forms but is more active on the lipid-modified form. In the present study, we compared more detailed kinetic properties of Smg GDS and mCdc25 by use of the lipid-modified form of Ki-Ras as a common substrate. Both Smg GDS and mCdc25 stimulated the dissociation of GDP from Ki-Ras and formed the stable binary complex with Ki-Ras. In the presence of guanosine 5'-(3-O-thio) triphosphate (GTP gamma S), the stable ternary complex of Smg GDS-GTP gamma S-Ki-Ras was produced, whereas GTP gamma S induced the dissociation of mCdc25 from mCdc25-Ki-Ras complex, yielding GTP gamma S-Ki-Ras. mCdc25 stimulated the dissociation of GDP from both the membrane-bound and soluble forms of Ki-Ras, whereas Smg GDS was far less active on the membrane-bound form than on the soluble form. Moreover, Smg GDS translocated the GTP gamma S-bound form of membrane-bound Ki-Ras to the soluble fraction as the stable ternary complex of Smg GDS-GTP gamma S-Ki-Ras, whereas mCdc25 did not show this activity. These results suggest that Smg GDS and mCdc25 play different roles in the regulation of Ki-Ras.
Nakanishi
H
H
Department of Biochemistry, Kobe University School of Medicine, Japan.
Kaibuchi
K
K
Orita
S
S
Ueno
N
N
Takai
Y
Y
D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D005656:Fungal Proteins / Q000378:metabolism*; D019204:GTP-Binding Proteins / Q000378:metabolism*; D006150:Guanine Nucleotides / Q000378:metabolism; D007700:Kinetics; D015689:Oncogene Protein p21(ras) / Q000378:metabolism; D010749:Phosphoprotein Phosphatases / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D020668:rap GTP-Binding Proteins; D020666:ras-GRF1
D005656:Fungal Proteins; D006150:Guanine Nucleotides; D020666:ras-GRF1; D010749:Phosphoprotein Phosphatases; D019204:GTP-Binding Proteins; D015689:Oncogene Protein p21(ras); D020668:rap GTP-Binding Proteins