A fast and efficient translational control system for conditional expression of yeast genes.
A new artificial regulatory system for essential genes in yeast is described. It prevents translation of target mRNAs upon tetracycline (tc) binding to aptamers introduced into their 5'UTRs. Exploiting direct RNA-ligand interaction renders auxiliary protein factors unnecessary. Therefore, our approach is strain independent and not susceptible to interferences by heterologous expressed regulatory proteins. We use a simple PCR-based strategy, which allows easy tagging of any target gene and the level of gene expression can be adjusted due to various tc aptamer-regulated promoters. As proof of concept, five differently expressed genes were targeted, two of which could not be regulated previously. In all cases, adding tc completely prevented growth and, as shown for Nop14p, rapidly abolished de novo protein synthesis providing a powerful tool for conditional regulation of yeast gene expression.
Kötter
Peter
P
Institut für Molekulare Biowissenschaften, Cluster of Excellence: Macromolecular Complexes, Johann Wolfgang Goethe-Universität Frankfurt, Max-von-Laue-Str. 9, D-60438 Frankfurt/M., Germany.
Weigand
Julia E
JE
Meyer
Britta
B
Entian
Karl-Dieter
KD
Suess
Beatrix
B
D020121:5' Untranslated Regions; D052157:Aptamers, Nucleotide / Q000737:chemistry; D015966:Gene Expression Regulation, Fungal; D020043:Genes, Essential; D005800:Genes, Fungal; D009687:Nuclear Proteins / Q000096:biosynthesis / Q000235:genetics; D016133:Polymerase Chain Reaction; D014176:Protein Biosynthesis; D012269:Ribosomal Proteins / Q000096:biosynthesis / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins / Q000096:biosynthesis / Q000235:genetics
D020121:5' Untranslated Regions; D052157:Aptamers, Nucleotide; C444458:NOP14 protein, S cerevisiae; D009687:Nuclear Proteins; D012269:Ribosomal Proteins; D029701:Saccharomyces cerevisiae Proteins