Post-translational regulation of Saccharomyces cerevisiae proteins tagged with the hormone-binding domains of mammalian nuclear receptors.
In the post-genome sequencing era the functional analysis of newly discovered proteins becomes more and more important. In this report we describe a genetic approach to the post-translational regulation of protein function in Saccharomyces cerevisiae by creating conditional lethal mutants. The yeast ORFs YDL139c, YDL147w, ERG3 and ERG11 were tagged with sequences encoding the hormone-binding domains of mammalian steroid receptors by PCR-mediated, targeted integration into the yeast genome. We found that the function of the chimeric proteins is regulated in a hormone-dependent way. This technique provides another important tool for the functional analysis of the yeast proteome.
Launhardt
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Department of Cell and Molecular Biology, Hans-Knöll-Institut für Naturstoff-Forschung, Jena, Germany.
Munder
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D000818:Animals; D002455:Cell Division; D002467:Cell Nucleus / Q000378:metabolism*; D006728:Hormones / Q000378:metabolism; D007700:Kinetics; D008024:Ligands; D008954:Models, Biological; D008957:Models, Genetic; D016296:Mutagenesis; D016366:Open Reading Frames; D010957:Plasmids / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D017434:Protein Structure, Tertiary; D011960:Receptors, Estrogen / Q000235:genetics; D011965:Receptors, Glucocorticoid / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism*; D013997:Time Factors; D014170:Transformation, Genetic
D006728:Hormones; D008024:Ligands; D011960:Receptors, Estrogen; D011965:Receptors, Glucocorticoid