Summary
The chorismate mutase structural gene, ARO7, which is necessary for both phenylalanine and tyrosine biosynthesis was cloned by complementation in yeast. Genetic analysis showed that ARO7 was identical to a gene necessary for growth in hypertonic medium, OSM2, which mapped nearby. After restriction mapping and subcloning of the plasmid, the cloned gene was used to detect mRNA levels in several growth conditions. Enzyme activities were measured in various genotypes. At our level of detection ARO7-OSM2 is a low level constitutively expressed gene.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Beggs JD (1978) Transformation of yeast by a replicating hybrid plasmid. Nature 275:104–109
Botstein D, Falco SC, Stewart M, Brennan M, Scherer S, Stinchcomb DT, Struhl K, Davis RW (1979) Sterile host yeasts (SHY): a cukaryotic system of biological containment for recombinant DNA experiments. Gene 8:17–24
Broach JR, Strathern JN, Hicks JB (1979) Transformation in yeast: development of a hybrid cloning vector and isolation of the Can1 gene. Gene 8:121–133
Conde J, Fink GR (1976) A mutant of Saccharomyces cerevisiae defective for nuclear fusion. Proc Natl Acad Sci USA 69:2846–2849
Hawthorne DC, Mortimer RK (1968) Genetic mapping of nonsense suppressors in yeast. Genetics 60:735–742
Jones EW, Fink GR (1982) Regulation of amino acid and nucleotide biosynthesis in yeast. In: Strathern JN, Jones EW, Broach JR (eds) The molecular biology of yeast saccharomyces: metabolism and gene expression. Cold Spring Harbor Laboratory, New York, pp 181–299
Kradolfer P, Zeyer J, Miozzari G, Huetter R (1977) Dominant regulatory mutants in chorismate mutase of Saccharomyces cerevisiae. FEMS Microbiol Lett 2:211
Lingens F, Goebel W, Uesseler H (1966) Regulation der Biosynthese der aromatischen Aminosäuren in Saccharomyces cerevisiae. 1. Hemmung der Enzymaktivitäten (Feedbackwirkung). Biochem Z 346:357–367
Lingens F, Goebel W, Uesseler H (1967) Regulation der aromatischen Aminosäuren in Saccharomyces cerevisiae. 2. Repression, Induktion, und Aktivierung. eur J Biochem 1:363–374
Lowry OH, Rosebrough NJ, Farr AC, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275
Maniatis T, Frisch E, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, New York
Meade JH, Manney TR (1983) Sensitivity of tryptophan, tyrosine, and phenylalanine mutants of Saccharomyces cerevisiae to phenethyalcohol. Genetics 104:235–240
Messenguy F, Dubois E (1983) Participation of transcriptional and post-transcriptional regulatory mechanisms in the control of arginine metabolism in yeast. Mol Gen Genet 189:148–156
Mortimer RK, Hawthorne DC (1975) Genetic mapping in yeast. Methods Cell Biol 11:221–233
Nasmyth KA, Reed SI (1980) Isolation of genes by complementation in yeast: molecular cloning of a cell cycle gene. Proc Natl Acad Sci USA 77:2119–2123
Singh A, Sherman F (1978) Deletions of the iso-1-cytochrome c and adjacent genes of yeast: discovery of the OSM1 gene controlling osmotic sensitivity. Genetics 89:653–655
Struhl K, Stinchcomb DT, Scherer S, Davis RW (1979) High-frequency transformation of yeast: autonomous replication of hybrid DNA molecules. Proc Natl Acad Sci USA 76:1035–1039
Wickner RB (1979) Mapping chromosomal genes of Saccharomyces cerevisiae using an improved genetic mapping method. Genetics 92:808–821
Wickner RB (1980) Plasmids controlling exclusion of the K2 killer double-stranded RNA plasmid of yeast. Cell 21:217–226
Wickner RB, Leibowitz MJ (1976) Chromosomal genes essential for replication of a double-stranded RNA plasmid of Saccharomyces cerevisiae: the killer character of yeast. J Mol Biol 105:427–443
Author information
Authors and Affiliations
Additional information
Communicated by C.P. Hollenberg
Rights and permissions
About this article
Cite this article
Ball, S.G., Wickner, R.B., Cottarel, G. et al. Molecular cloning and characterization of ARO7-OSM2, a single yeast gene necessary for chorismate mutase activity and growth in hypertonic medium. Molec. Gen. Genet. 205, 326–330 (1986). https://doi.org/10.1007/BF00430446
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00430446