Summary
Regulation of the two isozymes of 3-deoxy-D-arabino-heptulosonate-7-phosphate synthase (DAHP synthase; EC 4.1.2.15) encoded by the genes ARO3 and ARO4 of Saccharomyces cerevisiae was studied. Both genes were shown to respond equally well to the general control of amino acid biosynthesis. Strains with mutations in these two genes were obtained by selecting first for a single aro3 mutation and afterwards for a double aro3 aro4 mutation. Gene ARO3, coding for the phenylanine-dependent isozyme of DAHP synthase was cloned on the 2 μm multicopy vector pJDB207 by complementation of mutation aro3-1 in yeast. The ARO3 gene, carried originally on a 9.6 kb BamHI fragment (plasmid pME541A), was subcloned on a 1.9 kb HindIII-XbaI fragment (plasmid pME543). A transcript of about 1.5 kb was shown to proceed from the HindIII towards the XbaI site. Expression from the 9.6 kb as well as from the 1.9 kb fragment was normal on a multicopy vector, since in both cases DAHP synthase levels of about 50-fold the wild-type level were observed.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Aebi M, Niederberger P, Hütter R (1982) Isolation of the TRP2 and the TRP3 genes of Saccharomyces cerevisiae. Curr Genet 5:39–46
Aebi M, Furter R, Prantl F, Niederberger P, Hütter R (1984) Structure and function of the TRP3 gene of Saccharomyces cerevisiae: analysis of transcription, promoter sequence, and sequence coding for a glutamine amidotransferase. Curr Genet 8:165–172
Donahue TF, Daves RS, Lucchini G, Fink GR (1983) A short nucleotide sequence required for regulation of HIS4 by the general control system of yeast. Cell 32:89–98
Gollub E, Zalkin H, Sprinson DB (1970) Assay for 3-deoxy-D-arabino-heptulosonic acid 7-phosphate synthase. Methods Enzymol 17A:349–350
Herbert D, Phipps PJ, Strange RE (1971) Chemical analysis of microbial cells. Methods Microbiol 5B:209–240
Hope IA, Struhl K (1985) GCN4 protein, synthesized in vitro, binds HIS3 regulatory sequences: implications for general control of amino acid biosynthetic genes in yeast. Cell 43:177–188
Kieser T (1984) Factors affecting the isolation of CCC DNA from Streptomyces lividans and Escherichia coli. Plasmid 12:19–36
Lingens F, Goebel W, Uesseler H (1967) Regulation der biosynthese der aromatischen Aminosäuren in Saccharomyces cerevisiae. Eur J Biochem 1:363–374
Lucchini G, Hinnebusch AG, Chen C, Fink GR (1984) Positive regulatory interactions of the HIS4 gene of Saccharomyces cerevisiae. Mol Cell Biol 4:1326–1333
Melton DA, Krieg PA, Rebagliati MR, Maniatis T, Zinn K, Green MR (1984) Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acids Res 12:7035–7056
Messenguy F, Delforge J (1976) Role of transfer ribonucleic acids in the regulation of several biosyntheses in Saccharomyces cerevisiae. Eur J Biochem 67:335–339
Meuris P, Lacroute F, Slonimski PP (1967) Etude systématique de mutants inhibés par leurs propres métabolites chez la levure Saccharomyces cerevisiae. Genetics 56:149–161
Miller JH (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York,p 433
Miozzari G, Niederberger P, Hütter R (1978) Tryptophan biosynthesis in Saccharomyces cerevisiae: control of the flux through the pathway. J Bacteriol 134:48–59
Niederberger P, Miozzari G, Hütter R (1981) Biological role of the general control of amino acid biosynthesis in Saccharomyces cerevisiae. Mol Cell Biol 1:584–593
Schürch A, Miozzari G, Hütter R (1974) Regulation of tryptophan biosynthesis in Saccharomyces cerevisiae: mode of action of 5-methyltryptophan and 5-methyltryptophan-sensitive mutants. J Bacteriol 117:1131–1140
Sherman F, Fink GR, Lukins HB (1970) Methods in yeast genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, pp 51–53
Southern EM (1975) Detection of specific sequences among the DNA fragments separated by gel electrophoresis. J Mol Biol 98:503–517
Stäheli P, Agris PF, Niederberger P, Gehrke CW, Hütter R (1982) Accumulation of 2′-O-Methyl-guanosine deficient tRNATRP in tryptophan limited Saccharomyces cerevisiae. J Gen Microbiol 128:2591–2600
Takahashi M, Chan WWC (1971) Separation and properties of isozymes of 3-deoxy-D-arabino-heptulosonate-7-phosphate synthetase from Saccharomyces cerevisiae. Can J Biochem 49:1015–1025
Walton EF, Carter BLA, Pringle JR (1979) An enrichment method of temperature-sensitive and auxotrophic mutants of yeast. Mol Gen Genet 171:111–114
Zinn K, DiMaio D, Maniatis T (1983) Identification of two distinct regulatory regions adjacent to the human β-interferon gene. Cell 34:865–879
Author information
Authors and Affiliations
Additional information
Communicated by C.P. Hollenberg
Rights and permissions
About this article
Cite this article
Teshiba, S., Furter, R., Niederberger, P. et al. Cloning of the ARO3 gene of Saccharomyces cerevisiae and its regulation. Molec. Gen. Genet. 205, 353–357 (1986). https://doi.org/10.1007/BF00430450
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00430450