Nucleotide sequence of yeast LEU2 shows 5′-noncoding region has sequences cognate to leucine

Andreadis, Athena; Hsu, Yun-Pung P.; Kohlhaw, Gunter B.; Schimmel, Paul

doi:10.1016/0092-8674(82)90125-8

Cited by 207 publications

(80 citation statements)

References 34 publications

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“…DNA fragments were transferred to nitrocellulose (Millipore HAWP 00010 or to Schleicher and Schuell BA85) by Southern transfer in 20 • SSC (33). The probes used in this study are pC503 (HIS4) ( 11,13) and the 0.9 kb EcoRI-SalI fragment (39) of pC512 (LEU2) (1,2,7,10,15,34). Both genes originate from chromosome III in S. cerevisiae.…”

Section: Molecular Hybridization Analysismentioning

confidence: 99%

DNA sequence polymorphisms in the genus Saccharomyces IV. Homoeologous chromosomes III of Saccharomyces bayanus, S. carlsbergensis, and S. uvarum

Pedersen

1986

Carlsberg Res. Commun.

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Single chromosome III transfers have been accomplished from the two species S. bayanus and S. uvarum into genetically marked S. cerevisiae strains. Incompabilities between S. bayanus or S. uvarnm, and S. cerevisiae strains may account for the very few chromosome addition lines obtained. The preliminary genetic map for S. bayanus chromosome III is similar to the genetic map for a S. carlsbergensis chromosome lII from Danish lager strain BK2208. Recombination is absent between the transferred chromosomes and the auxotrophically marked chromosome from S. cerevisiae in the interval HIS4 to LEU2 but recombination does occur in the interval LEU2 to THR4. The mapped S. bayanus chromosome III carries HIS4 pattern III while the mapped chromosome III from the lager strain contains HIS4 pattern II. Transfer of a chromosome III from S. uvarum to S. cerevisiae has been recognised by restriction endonuclease fragment patterns, OFAGE chromosome separations and molecular hybridization, but a genetic map could not be constructed for an S. uvarum chromosome due to instability of the chromosome addition lines. The hypothesis is put forward that the S. carlsbergensis type strain and the lager strains are sibling species and species hybrids both produced by hybridization of an S. cerevisiae top fermenting strain and the bottom fermenting strain S. monacensis. The lager strains and the authentic type strain are designated S. carlsbergensis. From the electrophoretic karyotypes of type strains of Saecharomyces and molecular hybridization patterns it is deduced that S. bayanus, S. inusitatus, S. pastorianus and S. uvarum are closely related but distinct from the bottom fermenting strains. I N T R O D U C T I O NBase composition, DNA-RNA homology, DNA-DNA reassociation kinetics and restriction endonuclease fragment length polymorphisms have proven useful in the taxonomic evaluations of genera and species as well as in the diagnosis of closely related yeast strains (3,4,15,21,23,24,25,26,27,28,30,40). For taxonomic as well as breeding purposes the establishment of genetic linkage groups and their assignment to Abbreviations: OFAGE = Orthogonal field alternation gel electrophoresis; TBE = 0.089 M-Tris-borate, 0.089 M-boric acid, 0.002 M-EDTA; SSC = 0.15 M-NaC1, 15 mM-Na citrate.Springer-Verlag

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Section: Molecular Hybridization Analysismentioning

confidence: 99%

DNA sequence polymorphisms in the genus Saccharomyces IV. Homoeologous chromosomes III of Saccharomyces bayanus, S. carlsbergensis, and S. uvarum

Pedersen

1986

Carlsberg Res. Commun.

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“…In the CYCI and GAL1O genes, DNA sequences more than 100 nucleotides upstream (5') from the transcriptional initiation site are deemed important to the control of transcription. Similarly, upstream regulation sites have been recognized for genes encoding amino acid biosynthetic enzymes under the general control system in S. cerevisiae (1,7,13,30). We intend to examine the regulation of citrate synthase transcription and determine whether similar DNA sequence elements can be identified.…”

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confidence: 99%

Derepression of citrate synthase in Saccharomyces cerevisiae may occur at the level of transcription.

Hoosein¹,

Lewin²

1984

Mol. Cell. Biol.

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Pulse-chase labeling in whole cells and cell-free protein synthesis were l,ed to The level of mitochondrial enzymes is finely tuned to the energy demands of the cell. In bakers' yeast (Saccharomyces cerevisiae), the synthesis of respiratory chain components and enzymes of the citric acid cycle is repressed in the absence of oxygen or in the presence of glucose (9,18,24,25). The reverse of this process, the derepression of mitochondrial enzymes, is most easily monitored spectroscopically by the reappearance of cytochromes, but the synthesis of many achromatic mitochondrial enzymes is also stimulated by the removal of glucose from the culture medium or by the introduction of air to an anaerobic culture. Recent evidence suggests that synthesis of iso-1-cytochrome c in S. cereivisiae is regulated at the stage of transcription by the availability of heme (10, 30).We have studied the induction of citrate synthase (citrate oxaloacetate lyase, EC 4.1.3.7) during the growth of yeast cells in the presence of glucose. In Nelurospora crassa, this enzyme is made in the cytoplasm as a larger precursor which is processed to a Fmaller mature form as it is imported into the mitochondrion (12). Preliminary reports suggest that a similar mode of synthesis and transport occurs in S. cerevisiae (2). Like many other yeast mitochondrial proteins (21), citrate synthase is made primarily on "free" polyribosomes in the cytoplasm and is transported into the organelle independently of protein synthesis (28). Our interest in this enzyme was stimulated by the report (25) that citrate synthase activity can be derepressed in S. cerevisiae even in the presence of cycloheximide, an inhibitor of cytoplasmic protein synthesis. This result may reflect the presence of a pool of latent enzyme whose activity can be induced by the appropriate stimulus.We have begun by investigating whether the derepression of citrate synthase occurs at the level of transcription. We report that the amount of translatable RNA which encodes a precursor form of citrate synthase increases as the enzyme level is induced.MATERIALS AND METHODS S. cerevisiae strains and growth conditions. Two haploid S. cerevisiae strains were employed in this study. These were D273-1OB (MA To, ATCC 25657) and D273-10B/la, a [rho-] Purification of citrate synthase. Citrate synthase was purified from isolated S. cerevisiae mitochrondria by affinity elution chromatography (5, 19), using a modification of the method of Davies and Scopes (5). We replaced the carboxymethyl cellulose chromatography with a Biorex 70 column in the same buffer system. Nearly all of the loaded citrate synthase activity was bound to the column, and no activity could be measured in either of the wash fractions. Final specific activity of the isolated enzyme was greater than 1,400 U/mg of protein and represented a 150-fold purification over the crude lysate. Enzyme units are defined as micromoles of reduced coenzyme A formed per minute and were assayed by the method of Parvin (22).Antibodies to citrate synthase were ...

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“…Homoeologous genes in the two genomes are distinguishable by their restriction endonuclease fragment patterns. This is also true for the LEU2 genes, located on chromosomes III, encoding the 13-isopropylmalate dehydrogenase (1,2,9,11,23). The two LEU2 genes in lager yeast strains can be identified by their SalI diagnostic restriction fragment patterns (23).…”

Section: Introductionmentioning

confidence: 94%

DNA sequence polymorphisms in the genus Saccharomyces. V. Cloning and characterization of aLEU2 gene from S. carlsbergensis

Casey

Pedersen

1988

Carlsberg Res. Commun.

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Nucleotide sequence of yeast LEU2 shows 5′-noncoding region has sequences cognate to leucine

Cited by 207 publications

References 34 publications

DNA sequence polymorphisms in the genus Saccharomyces IV. Homoeologous chromosomes III of Saccharomyces bayanus, S. carlsbergensis, and S. uvarum

DNA sequence polymorphisms in the genus Saccharomyces IV. Homoeologous chromosomes III of Saccharomyces bayanus, S. carlsbergensis, and S. uvarum

Derepression of citrate synthase in Saccharomyces cerevisiae may occur at the level of transcription.

DNA sequence polymorphisms in the genus Saccharomyces. V. Cloning and characterization of aLEU2 gene from S. carlsbergensis

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