Cloning and transcriptional analysis of the ADE6 gene of Saccharomyces cerevisiae.
The Saccharomyces cerevisiae gene, ADE6, encoding 5'-phosphoribosylformyl glycinamidine synthetase (EC 6.3.5.3) has been cloned by complementation of an ade6 auxotroph. Transformation of ade6 mutants with ADE6-carrying centromeric plasmids restored normal, adenine-independent growth behavior in the recipients. Strains containing a disrupted ade6 allele were constructed and behaved as stable adenine auxotrophs. Southern transfer and genetic analyses of strains carrying a disrupted ade6 allele demonstrated that the cloned gene was ADE6 and not a suppressor. The cloned ADE6 DNA was mapped on the RAD2-proximal fragment of chromosome VII by hybridization on yeast chromosomes separated by pulsed-field gel electrophoresis. Northern-blot hybridization experiments show that the ADE6 region produces two different mRNA species of approx. 5 and 2 kb. Disappearance of the larger, but not the smaller, transcript is associated with ade6 mutations. A threefold repression in the amount of the 5-kb ADE6 mRNA is observed when growth medium is supplemented with exogenous adenine.
Gianì
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Istituto Biosintesi Vegetali C.N.R., Milan, Italy.
Manoni
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Breviario
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D000225:Adenine / Q000494:pharmacology; D015152:Blotting, Northern; D015139:Blotting, Southern; D019733:Carbon-Nitrogen Ligases with Glutamine as Amide-N-Donor; D003001:Cloning, Molecular; D016521:Electrophoresis, Gel, Pulsed-Field; D015966:Gene Expression Regulation, Fungal / Q000235:genetics; D005800:Genes, Fungal / Q000235:genetics*; D008025:Ligases / Q000235:genetics*; D010957:Plasmids / Q000235:genetics; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics; D014158:Transcription, Genetic / Q000235:genetics; D014170:Transformation, Genetic / Q000235:genetics
D008025:Ligases; D019733:Carbon-Nitrogen Ligases with Glutamine as Amide-N-Donor; C022385:phosphoribosylformylglycinamidine synthetase; D000225:Adenine