Tamanoi, F., Walsh, M., Kataoka, T., Wigler, M. H. (1984) A product of yeast RAS-2 gene is a guanine nucleotide binding protein. Proceedings of the National Academy of Sciences of the United States of America, 81 (22). pp. 6924-6928.
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Abstract
Saccharomyces cerevisiae contains 2 genes, RAS1 and RAS2, which show remarkable homology to mammalian ras genes. To characterize these gene products, the RAS2 gene in yeast was expressed using an inducible GAL10 promoter. After labeling with [35S]methionine and immunoprecipitating with a monoclonal antibody Y13-259, which reacts with p21 encoded by mammalian ras genes, a major band having an apparent MW of 41,000 was detected. This band was also identified in cell-free translation products of polyadenylylated RNA extracted from yeast cells grown in the presence of galactose. Crude extracts of cells expressing the RAS2 gene exhibited guanine nucleotide binding activity. This was detected by incubation with [3H]GDP followed by immunoprecipitation with the antibody Y13-259. The binding of labeled GDP was inhibited by a 20-fold excess of GDP, GTP, and, to a lesser extent, by UTP, a characteristic similar to that possessed by the mammalian ras proteins. The activity of the yeast protein differs from that of the mammalian proteins in its strong dependence on temperature. The guanine nucleotide binding activity provides an assay to purify the yeast protein.
| Item Type: | Paper |
|---|---|
| Subjects: | bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > genes, structure and function > genes: types > RAS organism description > yeast > Saccharomyces bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > genes, structure and function > gene expression bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > genes, structure and function > genes: types > oncogene |
| CSHL Authors: | |
| Communities: | CSHL labs > Wigler lab |
| Depositing User: | CSHL Librarian |
| Date: | 1984 |
| Date Deposited: | 06 Apr 2012 13:51 |
| Last Modified: | 30 Sep 2019 19:46 |
| Related URLs: | |
| URI: | https://repository.cshl.edu/id/eprint/26004 |
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