A novel in vivo assay for the analysis of protein-protein interaction.
The Ras Recruitment System (RRS) is a method for identification and isolation of protein-protein interaction. The method is based on translocation of cytoplasmic mammalian Ras protein to the inner leaflet of the plasma membrane through protein-protein interaction. The system is studied in a temperature-sensitive yeast strain where the yeast Ras guanyl nucleotide exchange factor is inactive at 36 degrees C. Protein-protein interaction results in cell growth at the restrictive temperature. We developed a gene reporter assay for the analysis of protein-protein interaction in mammalian cells. Ras activation in mammalian cells induces the mitogen-activated kinase cascade (MAPK), which can be monitored using Ras-dependent reporter genes. This greatly extends the usefulness of the system and provides a novel assay for protein-protein interaction in mammalian cells.
Maroun
M
M
Department of Molecular Genetics, Rappaport Family Institute for Research in the Medical Sciences and the B. Rappaport Faculty of Medicine, Technion-Israel Institute of Technology, Bat-Galim, Haifa, Israel.
Aronheim
A
A
D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D051380:GRB2 Adaptor Protein; D017930:Genes, Reporter; D006801:Humans; D020935:MAP Kinase Signaling System; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D025941:Protein Interaction Mapping / Q000379:methods*; D021381:Protein Transport; D016283:Proto-Oncogene Proteins p21(ras) / Q000032:analysis* / Q000378:metabolism; D020664:Son of Sevenless Proteins / Q000378:metabolism
D048868:Adaptor Proteins, Signal Transducing; D051380:GRB2 Adaptor Protein; C495948:GRB2 protein, human; D020664:Son of Sevenless Proteins; D019869:Phosphatidylinositol 3-Kinases; C501009:HRAS protein, human; D016283:Proto-Oncogene Proteins p21(ras)