Genetic co-regulation of galactose and melibiose utilization in Saccharomyces

Kew, Olen M.; Douglas, H. C.

doi:10.1128/jb.125.1.33-41.1976

Cited by 109 publications

(31 citation statements)

References 27 publications

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“…GFP production was 2.4-fold higher in this gal1 mutant strain. We verified previously reported data that galactose is a gratuitous inducer of the GAL promoter in a gal1 mutant strain [18][19][20][21]. Moreover, we also proved that in the gal1 mutant strain, protein production was significantly higher not only in comparison to the wild-type, but also in comparison to the other 27 deletion strains.…”

Section: Level Of Protein Production In Various Yeast Mutant Strainssupporting

confidence: 90%

Fluorescence based assay ofGALsystem in yeastSaccharomyces cerevisiae

Štagoj¹,

Comino²,

Komel³

2005

FEMS Microbiology Letters

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The GAL1 promoter is one of the strongest inducible promoters in the yeast Saccharomyces cerevisiae. In order to improve recombinant protein production we have developed a fluorescence based method for screening and evaluating the contribution of various gene deletions to protein expression from the GAL1 promoter. The level of protein synthesis was determined in 28 selected mutant strains simultaneously, by direct measurement of fluorescence in living cells using a microplate reader. The highest, 2.4-fold increase in GFP production was observed in a gal1 mutant strain. Deletion of GAL80 caused a 1.3-fold increase in fluorescence relative to the isogenic strain. GAL3, GAL4 and MTH1 gene deletion completely abrogated GFP synthesis. Growth of gal7, gal10 and gal3 also exhibited reduced fitness in galactose medium. Other genetic perturbations affected the GFP expression level only moderately. The fluorescence based method proved to be useful for screening genes involved in GAL1 promoter regulation and provides insight into more efficient manipulation of the GAL system.

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Section: Level Of Protein Production In Various Yeast Mutant Strainssupporting

confidence: 90%

Fluorescence based assay ofGALsystem in yeastSaccharomyces cerevisiae

Štagoj¹,

Comino²,

Komel³

2005

FEMS Microbiology Letters

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“…In vitro studies have demonstrated that dimerization of Gal80p stabilizes the Gal4p-Gal80p and the DNA-Gal4p-Gal80p complexes [7]. The GAL genes are expressed in presence of galactose through Gal3p, which relieves the inhibitory effect of Gal80p [20,22,24,[28][29][30].…”

Section: The Gal Genetic Switchmentioning

confidence: 99%

A steady‐state modeling approach to validate an in vivo mechanism of the GAL regulatory network in Saccharomyces cerevisiae

Verma¹,

Bhat

Bhartiya³

et al. 2004

European Journal of Biochemistry

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Cellular regulation is a result of complex interactions arising from DNA-protein and protein-protein binding, autoregulation, and compartmentalization and shuttling of regulatory proteins. Experiments in molecular biology have identified these mechanisms recruited by a regulatory network. Mathematical models may be used to complement the knowledge-base provided by in vitro experimental methods. Interactions identified by in vitro experiments can lead to the hypothesis of multiple candidate models explaining the in vivo mechanism. The equilibrium dissociation constants for the various interactions and the total component concentration constitute constraints on the candidate models. In this work, we identify the most plausible in vivo network by comparing the output response to the experimental data. We demonstrate the methodology using the GAL system of Saccharomyces cerevisiae for which the steady-state analysis reveals that Gal3p neither dimerizes nor shuttles between the cytoplasm and the nucleus.

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“… Predicted pattern of Gal gene expression as a function of the galactose concentration in a wild‐type strain. The symbols represent experimental data as reported by Kew and Douglas (1976). ★ denotes transferase, and · denotes α‐galactosidase activities.…”

Section: Resultsmentioning

confidence: 99%

Quantitative Model for Gal4p-Mediated Expression of the Galactose/Melibiose Regulon in Saccharomyces cerevisiae

et al. 1999

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A mathematical model based on equilibrium binding between Gal4p and its specific DNA binding site has been developed. A model for GAL gene expression solely due to cooperativity, as a function of Gal4p concentration, has been developed for a gal80 mutant. The above model was extended to include other known regulatory molecules, namely Gal80p and Gal3p. Parameters determined from the above simulation were then used to represent a physiological status of gene expression in response to glucose (in terms of Gal4p concentration) and galactose in a wild-type strain. We demonstrate that in a wild-type strain glucose repression is more stringent due to cooperativity and autogenous regulation, while the induction response to galactose is only through autogenous regulation. The biological significance of autogenous regulation in Saccharomyces cerevisiae is discussed vis-a-vis the lactose operon of Escherichia coli.

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Genetic co-regulation of galactose and melibiose utilization in Saccharomyces

Cited by 109 publications

References 27 publications

Fluorescence based assay ofGALsystem in yeastSaccharomyces cerevisiae

Fluorescence based assay ofGALsystem in yeastSaccharomyces cerevisiae

A steady‐state modeling approach to validate an in vivo mechanism of the GAL regulatory network in Saccharomyces cerevisiae

Quantitative Model for Gal4p-Mediated Expression of the Galactose/Melibiose Regulon in Saccharomyces cerevisiae

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