| Title: | The N-terminal non-RGS domain of human regulator of G-protein signalling 1 contributes to its ability to inhibit pheromone receptor signalling in yeast |
| Author(s): | Somerville W; Song W; Kong JL; Panetta R; Greenwood MT; |
| Address: | "Department of Anatomy and Cell Biology, McGill University, Montreal, Quebec, Canada" |
| DOI: | 10.1016/s0898-6568(02)00121-3 |
| ISSN/ISBN: | 0898-6568 (Print) 0898-6568 (Linking) |
| Abstract: | "Regulators of G-protein signalling (RGS) are a family of proteins that interact with G-proteins to regulate negatively G-protein coupled receptor (GPCR) signalling. In addition to a conserved core domain that is necessary and sufficient for their GTPase activating protein (GAP) like activity, RGSs possess N- and C-terminal motifs that confer distinct functional differences. In order to identify the role of the non-RGS region of human RGS1, we have characterized a series of fusions between RGS1 and GFP in a yeast mutant lacking the RGS containing SST2 gene. Using both halo assays as well as a GPCR responsive FUS1-LacZ reporter gene, we demonstrate that a RGS1-GFP fusion inhibits GPCR signalling in yeast while GFP fusions containing either the N-terminus non RGS sequence of RGS1(1-68) or the sequence containing the RGS box of RGS1(68-197) produce proteins that retain RGS1 activity. These results suggest that both the N-terminal and the RGS box of RGS1 function to inhibit signalling. Analysis of a series of mutants spanning the entire N-terminal non-RGS region of RGS1 produced by conservative segment exchange (CSE) mutagenesis showed little loss of function in yeast. This suggests that the overall structure of the N-terminal region of RGS1 rather than specific motifs or residues is required for its function" |
| Keywords: | "Amino Acid Sequence GTP-Binding Protein Regulators/*genetics/metabolism GTP-Binding Proteins/*metabolism GTPase-Activating Proteins/genetics Genes, Reporter Humans Molecular Sequence Data Mutation Pheromones/*metabolism Plasmids Saccharomyces cerevisiae/*;" |
| Notes: | "MedlineSomerville, Wendy Song, Wei Kong, Janice L Panetta, Rosemarie Greenwood, Michael T eng Research Support, Non-U.S. Gov't England 2003/03/06 Cell Signal. 2003 Apr; 15(4):413-21. doi: 10.1016/s0898-6568(02)00121-3" |
