Activation of Transcription by Metabolic Intermediates of the Pyrimidine Biosynthetic Pathway

Overview

Journal Mol Cell Biol

Specialty Cell Biology

Date 1998 Dec 22

PMID 9858611

Citations 27

Authors

P J Flynn

R J Reece

Affiliations

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Abstract

Saccharomyces cerevisiae responds to pyrimidine starvation by increasing the expression of four URA genes, encoding the enzymes of de novo pyrimidine biosynthesis, three- to eightfold. The increase in gene expression is dependent on a transcriptional activator protein, Ppr1p. Here, we investigate the mechanism by which the transcriptional activity of Ppr1p responds to the level of pyrimidine biosynthetic intermediates. We find that purified Ppr1p is unable to promote activation of transcription in an in vitro system. Transcriptional activation by Ppr1p can be observed, however, if either dihydroorotic acid (DHO) or orotic acid (OA) is included in the transcription reactions. The transcriptional activation function and the DHO/OA-responsive element of Ppr1p localize to the carboxyl-terminal 134 amino acids of the protein. Thus, Ppr1p directly senses the level of early pyrimidine biosynthetic intermediates within the cell and activates the expression of genes encoding proteins required later in the pathway. These results are discussed in terms of (i) regulation of the pyrimidine biosynthetic pathway and (ii) a novel mechanism of regulating gene expression.

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References

Friden P, Schimmel P . LEU3 of Saccharomyces cerevisiae activates multiple genes for branched-chain amino acid biosynthesis by binding to a common decanucleotide core sequence. Mol Cell Biol. 1988; 8(7):2690-7. PMC: 363479. DOI: 10.1128/mcb.8.7.2690-2697.1988. View

Loison G, Losson R, Lacroute F . Constitutive mutants for orotidine 5 phosphate decarboxylase and dihydroorotic acid dehydrogenase in Saccharomyces cerevisiae. Curr Genet. 2013; 2(1):39-44. DOI: 10.1007/BF00445692. View

Sze J, Woontner M, Jaehning J, Kohlhaw G . In vitro transcriptional activation by a metabolic intermediate: activation by Leu3 depends on alpha-isopropylmalate. Science. 1992; 258(5085):1143-5. DOI: 10.1126/science.1439822. View

Ohashi Y, Brickman J, Furman E, Middleton B, Carey M . Modulating the potency of an activator in a yeast in vitro transcription system. Mol Cell Biol. 1994; 14(4):2731-9. PMC: 358639. DOI: 10.1128/mcb.14.4.2731-2739.1994. View

Friden P, Schimmel P . LEU3 of Saccharomyces cerevisiae encodes a factor for control of RNA levels of a group of leucine-specific genes. Mol Cell Biol. 1987; 7(8):2708-17. PMC: 367887. DOI: 10.1128/mcb.7.8.2708-2717.1987. View

Lacroute F . Regulation of pyrimidine biosynthesis in Saccharomyces cerevisiae. J Bacteriol. 1968; 95(3):824-32. PMC: 252099. DOI: 10.1128/jb.95.3.824-832.1968. View

Liang S, Marmorstein R, Harrison S, Ptashne M . DNA sequence preferences of GAL4 and PPR1: how a subset of Zn2 Cys6 binuclear cluster proteins recognizes DNA. Mol Cell Biol. 1996; 16(7):3773-80. PMC: 231373. DOI: 10.1128/MCB.16.7.3773. View

Antonelli R, Estevez L . Carbamyl-phosphate synthetase domain of the yeast multifunctional protein Ura2 is necessary for aspartate transcarbamylase inhibition by UTP. FEBS Lett. 1998; 422(2):170-4. DOI: 10.1016/s0014-5793(98)00010-6. View

Friden P, Reynolds C, Schimmel P . A large internal deletion converts yeast LEU3 to a constitutive transcriptional activator. Mol Cell Biol. 1989; 9(9):4056-60. PMC: 362471. DOI: 10.1128/mcb.9.9.4056-4060.1989. View

10.

Ponticelli A, Struhl K . Analysis of Saccharomyces cerevisiae his3 transcription in vitro: biochemical support for multiple mechanisms of transcription. Mol Cell Biol. 1990; 10(6):2832-9. PMC: 360644. DOI: 10.1128/mcb.10.6.2832-2839.1990. View

11.

Jaquet L, Serre V, Lollier M, Penverne B, Herve G, Souciet J . Allosteric regulation of carbamoylphosphate synthetase-aspartate transcarbamylase multifunctional protein of Saccharomyces cerevisiae: selection, mapping and identification of missense mutations define three regions involved in feedback inhibition.... J Mol Biol. 1995; 248(3):639-52. DOI: 10.1006/jmbi.1995.0248. View

12.

Losson R, Lacroute F . Cloning of a eukaryotic regulatory gene. Mol Gen Genet. 1981; 184(3):394-9. DOI: 10.1007/BF00352511. View

13.

Schjerling P, Holmberg S . Comparative amino acid sequence analysis of the C6 zinc cluster family of transcriptional regulators. Nucleic Acids Res. 1996; 24(23):4599-607. PMC: 146297. DOI: 10.1093/nar/24.23.4599. View

14.

De Montigny J, Kern L, Hubert J, Lacroute F . Cloning and sequencing of URA10, a second gene encoding orotate phosphoribosyl transferase in Saccharomyces cerevisiae. Curr Genet. 1990; 17(2):105-11. DOI: 10.1007/BF00312853. View

15.

Potier S, Lacroute F, Hubert J, Souciet J . Studies on transcription of the yeast URA2 gene. FEMS Microbiol Lett. 1990; 60(1-2):215-9. DOI: 10.1016/0378-1097(90)90374-y. View

16.

Roy A, Exinger F, Losson R . cis- and trans-acting regulatory elements of the yeast URA3 promoter. Mol Cell Biol. 1990; 10(10):5257-70. PMC: 361211. DOI: 10.1128/mcb.10.10.5257-5270.1990. View

17.

Liljelund P, Losson R, Kammerer B, Lacroute F . Yeast regulatory gene PPR1. II. Chromosomal localization, meiotic map, suppressibility, dominance/recessivity and dosage effect. J Mol Biol. 1984; 180(2):251-65. DOI: 10.1016/s0022-2836(84)80003-0. View

18.

Marmorstein R, Harrison S . Crystal structure of a PPR1-DNA complex: DNA recognition by proteins containing a Zn2Cys6 binuclear cluster. Genes Dev. 1994; 8(20):2504-12. DOI: 10.1101/gad.8.20.2504. View

19.

Reece R, Rickles R, Ptashne M . Overproduction and single-step purification of GAL4 fusion proteins from Escherichia coli. Gene. 1993; 126(1):105-7. DOI: 10.1016/0378-1119(93)90596-u. View

20.

. Pyrimidine biosynthesis in Saccharomyces cerevisiae: the ura2 cluster gene, its multifunctional enzyme product, and other structural or regulatory genes involved in de novo UMP synthesis. Biochem Cell Biol. 1989; 67(9):612-31. DOI: 10.1139/o89-094. View