Analysis of interaction between Ras and CDC25 guanine nucleotide exchange factor using yeast GAL4 two-hybrid system.
Our results demonstrate that the GAL4 two-hybrid system can be useful for studying interactions of the wild-type and mutant forms of Ras proteins with the CDC25 guanine nucleotide exchange factor (CDC25-GEF). In addition, our findings show that a negative result in the GAL4 two-hybrid system does not indicate that the two proteins tested do not interact under all conditions but only that they do not interact under the specific conditions examined. We recommend that the two-hybrid system be employed in combination with other approaches, including molecular genetic analyses and in vitro binding experiments, for the study of Ras and CDC25-GEF interactions.
Department of Biochemistry and Molecular Biology, Norris Comprehensive Cancer Center, University of Southern California School of Medicine, Los Angeles 90033, USA.
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D000595:Amino Acid Sequence; D001483:Base Sequence; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism*; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D004268:DNA-Binding Proteins; D004926:Escherichia coli / Q000235:genetics; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D005822:Genetic Vectors; D006801:Humans; D066298:In Vitro Techniques; D008969:Molecular Sequence Data; D010749:Phosphoprotein Phosphatases / Q000235:genetics / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D014170:Transformation, Genetic; D018631:ras Proteins / Q000235:genetics / Q000378:metabolism*; D020666:ras-GRF1
D018797:Cell Cycle Proteins; D018076:DNA, Complementary; D004268:DNA-Binding Proteins; D005656:Fungal Proteins; C052130:GAL4 protein, S cerevisiae; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D020666:ras-GRF1; D010749:Phosphoprotein Phosphatases; D018631:ras Proteins