Zoller, M. J., Yonemoto, W., Taylor, S. S., Johnson, K. E. (March 1991) Mammalian cAMP-dependent protein kinase functionally replaces its homolog in yeast. Gene, 99 (2). pp. 171-9. ISSN 0378-1119 (Print)0378-1119 (Linking)
Abstract
The cDNA encoding the catalytic subunit (C alpha) from mouse cAMP-dependent protein kinase (PK) was expressed in Saccharomyces cerevisiae. By a plasmid swap procedure, we demonstrated that the mammalian C alpha subunit can functionally replace its yeast homolog to maintain the viability of a yeast strain containing genetic disruptions of the three TPK genes encoding the yeast C subunits. C alpha subunit produced in yeast was purified and its biochemical properties were determined. The protein isolated from yeast appears to be myristylated, as has been found for C subunits from higher eukaryotic cells. This system would be useful for studying the biochemistry of the mammalian enzyme in vitro and its biological role in a model in vivo system. These studies demonstrate that the PK substrate(s) required for viability are recognized by the mammalian enzyme. In general terms, these results demonstrate that heterologous proteins with only 50% sequence conservation with their yeast counterparts can be functional in yeast. This is an important result because it validates the use of yeast to identify the biological role of newly cloned genes from heterologous systems, a key tenet of the Human Genome Initiative.
| Item Type: | Paper |
|---|---|
| Uncontrolled Keywords: | Amino Acid Sequence Animals Base Sequence Chromosome Mapping Mice Molecular Sequence Data Plasmids Protein Kinases/*genetics/metabolism Recombination, Genetic Saccharomyces cerevisiae/genetics |
| Subjects: | bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > transcription factor > Cyclic AMP bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > enzymes > kinase organism description > yeast |
| CSHL Authors: | |
| Communities: | CSHL labs > Wigler lab |
| Depositing User: | Matt Covey |
| Date: | 15 March 1991 |
| Date Deposited: | 04 Jan 2016 23:02 |
| Last Modified: | 04 Jan 2016 23:02 |
| Related URLs: | |
| URI: | https://repository.cshl.edu/id/eprint/32104 |
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