scholarly journals Constitutive expression in gal7 mutants of Kluyveromyces lactis is due to internal production of galactose as an inducer of the Gal/Lac regulon.

1997 ◽  
Vol 17 (3) ◽  
pp. 1722-1730 ◽  
Author(s):  
G Cardinali ◽  
V Vollenbroich ◽  
M S Jeon ◽  
A A de Graaf ◽  
C P Hollenberg
Keyword(s):  
Culture Medium ◽  
Kluyveromyces Lactis ◽  
Constitutive Expression ◽  
Regulatory Function ◽  
Nuclear Magnetic Resonance Analysis ◽  
Resonance Analysis ◽  
Crude Extracts ◽  
Induction Process ◽  
Beta Galactosidase ◽  
Internal Production

The induction process of the galactose regulon has been intensively studied, but until now the nature of the inducer has remained unknown. We have analyzed a delta gal7 mutant of the yeast Kluyveromyces lactis, which lacks the galactotransferase activity and is able to express the genes of the Gal/Lac regulon also in the absence of galactose. We found that this expression is semiconstitutive and undergoes a strong induction during the stationary phase. The gal1-209 mutant, which has a reduced kinase activity but retains its positive regulatory function, also shows a constitutive expression of beta-galactosidase, suggesting that galactose is the inducer. A gal10 deletion in delta gal7 or gal1-209 mutants reduces the expression to under wild-type levels. The presence of the inducer could be demonstrated in both delta gal7 crude extracts and culture medium by means of a bioassay using the induction in gal1-209 cells. A mutation in the transporter gene LAC12 decreases the level of induction in gal7 cells, indicating that galactose is partly released into the medium and then retransported into the cells. Nuclear magnetic resonance analysis of crude extracts from delta gal7 cells revealed the presence of 50 microM galactose. We conclude that galactose is the inducer of the Gal/Lac regulon and is produced via UDP-galactose through a yet-unknown pathway.

Galactokinase encoded by GAL1 is a bifunctional protein required for induction of the GAL genes in Kluyveromyces lactis and is able to suppress the gal3 phenotype in Saccharomyces cerevisiae

1991 ◽  
Vol 11 (11) ◽  
pp. 5454-5461
Author(s):  
J Meyer ◽  
A Walker-Jonah ◽  
C P Hollenberg
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Kluyveromyces Lactis ◽  
Regulatory Function ◽  
Bifunctional Protein ◽  
Leloir Pathway ◽  
Gal Genes ◽  
Beta Galactosidase ◽  
Gal1 Gene ◽  
Galactokinase Activity ◽  
Negative Mutant

We have analyzed a GAL1 mutant (gal1-r strain) of the yeast Kluyveromyces lactis which lacks the induction of beta-galactosidase and the enzymes of the Leloir pathway in the presence of galactose. The data show that the K. lactis GAL1 gene product has, in addition to galactokinase activity, a function required for induction of the lactose system. This regulatory function is not dependent on galactokinase activity, as it is still present in a galactokinase-negative mutant (gal1-209). Complementation studies in Saccharomyces cervisiae show that K. lactis GAL1 and gal1-209, but not gal1-r, complement the gal3 mutation. We conclude that the regulatory function of GAL1 in K. lactis soon after induction is similar to the function of GAL3 in S. cerevisiae.

scholarly journals Galactokinase encoded by GAL1 is a bifunctional protein required for induction of the GAL genes in Kluyveromyces lactis and is able to suppress the gal3 phenotype in Saccharomyces cerevisiae.

1991 ◽  
Vol 11 (11) ◽  
pp. 5454-5461 ◽  
Author(s):  
J Meyer ◽  
A Walker-Jonah ◽  
C P Hollenberg
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Kluyveromyces Lactis ◽  
Regulatory Function ◽  
Bifunctional Protein ◽  
Leloir Pathway ◽  
Gal Genes ◽  
Beta Galactosidase ◽  
Gal1 Gene ◽  
Galactokinase Activity ◽  
Negative Mutant

We have analyzed a GAL1 mutant (gal1-r strain) of the yeast Kluyveromyces lactis which lacks the induction of beta-galactosidase and the enzymes of the Leloir pathway in the presence of galactose. The data show that the K. lactis GAL1 gene product has, in addition to galactokinase activity, a function required for induction of the lactose system. This regulatory function is not dependent on galactokinase activity, as it is still present in a galactokinase-negative mutant (gal1-209). Complementation studies in Saccharomyces cervisiae show that K. lactis GAL1 and gal1-209, but not gal1-r, complement the gal3 mutation. We conclude that the regulatory function of GAL1 in K. lactis soon after induction is similar to the function of GAL3 in S. cerevisiae.

scholarly journals Glycine Betaine: Reserve Form of Choline inPenicillium fellutanum in Low-Sulfate Medium

1999 ◽  
Vol 65 (3) ◽  
pp. 1340-1342 ◽  
Author(s):  
Yong-Il Park ◽  
Marian L. Buszko ◽  
John E. Gander
Keyword(s):  
Nuclear Magnetic Resonance ◽  
Magnetic Resonance ◽  
Glycine Betaine ◽  
Culture Medium ◽  
Multiple Roles ◽  
Nuclear Magnetic Resonance Analysis ◽  
Fungal Metabolism ◽  
Resonance Analysis ◽  
Sulfate Medium ◽  
Nuclear Magnetic

ABSTRACT In spite of choline’s importance in fungal metabolism, its sources in cytoplasm have not been fully established. 13C nuclear magnetic resonance analysis of mycelial extracts from day-5Penicillium fellutanum cultures showed that, as well as choline-O-sulfate, intracellular glycine betaine is another reserve form of choline, depending on the availability of sulfate in the culture medium. These observations are discussed relative to the multiple roles of choline and its precursors in P. fellutanum.

scholarly journals Tunneling Spectroscopy by Nuclear Magnetic Resonance: Analysis of Rotational Tunneling in Solid Pentamethylbenzene

1985 ◽  
Vol 40 (11) ◽  
pp. 1075-1084
Author(s):  
W. T. Sobol ◽  
K.R. Sridharan ◽  
I. G. Cameron ◽  
M. M. Pintar
Keyword(s):  
Frequency Dependence ◽  
Lattice Relaxation ◽  
Time Correlation ◽  
Lattice Relaxation Time ◽  
Polarization Transfer ◽  
Spin Lattice Relaxation ◽  
Transfer Model ◽  
Nuclear Magnetic Resonance Analysis ◽  
Spin Lattice ◽  
Resonance Analysis

The frequency dependence of the spin-lattice relaxation time T1 was measured at three temperatures near one of the Zeeman-tunneling level matching resonances for pentamethylbenzene. These measurements are correlated with 71 temperature dependence data from the literature. It is shown that the frequency dependence of the Zeeman-torsion coupling time cannot be explained in terms of the semiclassical perturbation theory using time correlation functions. A three bath polarization transfer model is also employed and the applicability of both models discussed. Zeeman-torsion coupling is further investigated using a saturation sequence and the results are compared with the predictions of the three bath polarization transfer model.

scholarly journals Functional homology between the yeast regulatory proteins GAL4 and LAC9: LAC9-mediated transcriptional activation in Kluyveromyces lactis involves protein binding to a regulatory sequence homologous to the GAL4 protein-binding site.

1987 ◽  
Vol 7 (12) ◽  
pp. 4400-4406 ◽  
Author(s):  
K D Breunig ◽  
P Kuger
Keyword(s):  
Protein Binding ◽  
Binding Site ◽  
Transcriptional Activation ◽  
Kluyveromyces Lactis ◽  
Regulatory Protein ◽  
Binding Activity ◽  
Protein Binding Site ◽  
Regulatory Sequence ◽  
Functional Homology ◽  
Beta Galactosidase

As shown previously, the beta-galactosidase gene of Kluyveromyces lactis is transcriptionally regulated via an upstream activation site (UASL) which contains a sequence homologous to the GAL4 protein-binding site in Saccharomyces cerevisiae (M. Ruzzi, K.D. Breunig, A.G. Ficca, and C.P. Hollenberg, Mol. Cell. Biol. 7:991-997, 1987). Here we demonstrate that the region of homology specifically binds a K. lactis regulatory protein. The binding activity was detectable in protein extracts from wild-type cells enriched for DNA-binding proteins by heparin affinity chromatography. These extracts could be used directly for DNase I and exonuclease III protection experiments. A lac9 deletion strain, which fails to induce the beta-galactosidase gene, did not contain the binding factor. The homology of LAC9 protein with GAL4 (J.M. Salmeron and S. A. Johnston, Nucleic Acids Res. 14:7767-7781, 1986) strongly suggests that LAC9 protein binds directly to UASL and plays a role similar to that of GAL4 in regulating transcription.

Polymyrcene microstructure revisited from precise high-field nuclear magnetic resonance analysis

Polymer ◽  
2014 ◽  
Vol 55 (16) ◽  
pp. 3869-3878 ◽  
Author(s):  
Sébastien Georges ◽  
Marc Bria ◽  
Philippe Zinck ◽  
Marc Visseaux
Keyword(s):  
Nuclear Magnetic Resonance ◽  
Magnetic Resonance ◽  
High Field ◽  
Nuclear Magnetic Resonance Analysis ◽  
Resonance Analysis ◽  
Nuclear Magnetic

Genetic studies on the role of the nucleoside transport function in nucleoside efflux, the inosine cycle, and purine biosynthesis

1983 ◽  
Vol 3 (7) ◽  
pp. 1187-1196
Author(s):  
B Ullman ◽  
K Kaur ◽  
T Watts
Keyword(s):  
Culture Medium ◽  
Purine Biosynthesis ◽  
Nucleoside Transport ◽  
Regulatory Function ◽  
Synthetase Activity ◽  
Common Component ◽  
Adenylosuccinate Synthetase ◽  
T Lymphoma Cells ◽  
T Lymphoma ◽  
Hypoxanthine Guanine Phosphoribosyltransferase

A mutant clone (AU-100) which is 90% deficient in adenylosuccinate synthetase activity was characterized from wild-type murine S49 T-lymphoma cells. This AU-100 cell line and its hypoxanthine-guanine phosphoribosyltransferase-deficient derivative, AUTG-50B, overproduce purines severalfold and excrete massive amounts of inosine into the culture medium (Ullman et al., Proc. Natl. Acad. Sci. U.S.A. 79:5127-5131, 1982). We introduced a mutation into both of these cell lines which make them incapable of taking up nucleosides from the culture medium. The genetic deficiency in nucleoside transport prevents the adenylosuccinate synthetase-deficient AU-100 cells from excreting inosine. Because of an extremely efficient intracellular inosine salvage system, the nucleoside transport-deficient AU-100 cells also no longer overproduce purines. AUTG-50B cells which have been made genetically deficient in nucleoside transport still overproduce purines but excrete hypoxanthine rather than inosine. These studies demonstrate genetically that nucleoside transport and nucleoside efflux share a common component and that nucleoside transport has an important regulatory function which profoundly affects the rates of purine biosynthesis and purine salvage.

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