Functional expression of mammalian cytochromes P450IIB in the yeast Saccharomyces cerevisiae

1991 ◽  
Vol 291 (1) ◽  
pp. 176-186 ◽  
Author(s):  
Karen M. Kedzie ◽  
Richard M. Philpot ◽  
James R. Halpert
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Functional Expression ◽  
Yeast Saccharomyces Cerevisiae

scholarly journals High-affinity iron and calcium transport pathways are involved in U(VI) uptake in the budding yeast Saccharomyces cerevisiae

2021 ◽  
Author(s):  
Benoît REVEL ◽  
Patrice CATTY ◽  
Stéphane RAVANEL ◽  
Jacques BOURGUIGNON ◽  
Claude ALBAN
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Living Organism ◽  
Functional Expression ◽  
Model Systems ◽  
Rapid Screening ◽  
Unicellular Eukaryote ◽  
Yeast Saccharomyces Cerevisiae ◽  
Transport Pathways ◽  
Living Organisms ◽  
Calcium Iron

Uranium (U) is a naturally-occurring radionuclide toxic for living organisms that can take it up. To date, the mechanisms of U uptake are far from being understood. Here, we used the yeast Saccharomyces cerevisiae as a unicellular eukaryote model to identify U assimilation pathways. Thus, we have identified, for the first time, transport machineries capable of transporting U in a living organism. First, we evidenced a metabolism-dependent U transport in yeast. Then, competition experiments with essential metals allowed us to identify calcium, iron and copper entry pathways as potential routes for U uptake. The analysis of various metal transport mutants revealed that mid1Δ, cch1Δ and ftr1Δ mutants, affected in calcium (Mid1/Cch1 channel) and Fe(III) (Ftr1/Fet3 complex) transport, respectively, exhibited highly reduced U uptake rates and accumulation, demonstrating the implication of these import systems in U uptake. Finally, expression of the Mid1 gene into the mid1Δ mutant restored U uptake levels of the wild type strain, underscoring the central role of the Mid1/Cch1 calcium channel in U absorption process in yeast. Our results also open up the opportunity for rapid screening of U-transporter candidates by functional expression in yeast, before their validation in more complex higher eukaryote model systems.

Functional expression of the cre-lox site-specific recombination system in the yeast Saccharomyces cerevisiae

1987 ◽  
Vol 7 (6) ◽  
pp. 2087-2096
Author(s):  
B Sauer
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Functional Expression ◽  
Yeast Saccharomyces Cerevisiae ◽  
Efficient Manner ◽  
Site Specific ◽  
Site Specific Recombination ◽  
Leu2 Gene ◽  
Recombination System ◽  
A Site ◽  
Site Specific Recombination System

The procaryotic cre-lox site-specific recombination system of coliphage P1 was shown to function in an efficient manner in a eucaryote, the yeast Saccharomyces cerevisiae. The cre gene, which codes for a site-specific recombinase, was placed under control of the yeast GALI promoter. lox sites flanking the LEU2 gene were integrated into two different chromosomes in both orientations. Excisive recombination at the lox sites (as measured by loss of the LEU2 gene) was promoted efficiently and accurately by the Cre protein and was dependent upon induction by galactose. These results demonstrate that a procaryotic recombinase can enter a eucaryotic nucleus and, moreover, that the ability of the Cre recombinase to perform precise recombination events on the chromosomes of S. cerevisiae is unimpaired by chromatin structure.

scholarly journals Functional expression of the cre-lox site-specific recombination system in the yeast Saccharomyces cerevisiae.

1987 ◽  
Vol 7 (6) ◽  
pp. 2087-2096 ◽  
Author(s):  
B Sauer
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Functional Expression ◽  
Yeast Saccharomyces Cerevisiae ◽  
Efficient Manner ◽  
Site Specific ◽  
Site Specific Recombination ◽  
Leu2 Gene ◽  
Recombination System ◽  
A Site ◽  
Site Specific Recombination System

The procaryotic cre-lox site-specific recombination system of coliphage P1 was shown to function in an efficient manner in a eucaryote, the yeast Saccharomyces cerevisiae. The cre gene, which codes for a site-specific recombinase, was placed under control of the yeast GALI promoter. lox sites flanking the LEU2 gene were integrated into two different chromosomes in both orientations. Excisive recombination at the lox sites (as measured by loss of the LEU2 gene) was promoted efficiently and accurately by the Cre protein and was dependent upon induction by galactose. These results demonstrate that a procaryotic recombinase can enter a eucaryotic nucleus and, moreover, that the ability of the Cre recombinase to perform precise recombination events on the chromosomes of S. cerevisiae is unimpaired by chromatin structure.

Functional Expression of Spirulina-Δ6 Desaturase Gene in Yeast, Saccharomyces cerevisiae

2005 ◽  
Vol 32 (4) ◽  
pp. 215-226 ◽  
Author(s):  
Pavinee Kurdrid ◽  
Sanjukta Subudhi ◽  
Apiradee Hongsthong ◽  
Marasri Ruengjitchatchawalya ◽  
Morakot Tanticharoen
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Functional Expression ◽  
Desaturase Gene ◽  
Yeast Saccharomyces Cerevisiae ◽  
Δ6 Desaturase

scholarly journals The functioning of mammalian ClC-2 chloride channel in Saccharomyces cerevisiae cells requires an increased level of Kha1p

2005 ◽  
Vol 390 (3) ◽  
pp. 655-664 ◽  
Author(s):  
Krzysztof Flis ◽  
Alexandre Hinzpeter ◽  
Aleksander Edelman ◽  
Anna Kurlandzka
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Golgi Apparatus ◽  
Chloride Channel ◽  
Mammalian Cells ◽  
Chloride Channels ◽  
Functional Expression ◽  
Ph Control ◽  
Gene Encoding ◽  
General Applicability ◽  
Yeast Saccharomyces Cerevisiae

The mammalian chloride channel ClC-2 is a member of the CLC voltage-gated chloride channels family. This broadly expressed protein shows diverse cellular locations and despite numerous studies, its precise function is poorly understood. Disruption of ClC-2-encoding gene in mouse leads to retinal and testicular degeneration and mutations in CLC2 (gene encoding the ClC-2 channel) are associated with idiopathic generalized epilepsies. ClC-2 may also be responsible for Cl− transport in mouse salivary glands. The only CLC homologue of the yeast Saccharomyces cerevisiae, Gef1p, exhibits CLC activity. We expressed the mammalian ClC-2 protein in S. cerevisiae devoid of Gef1p in an attempt to identify yeast proteins influencing the functioning of ClC-2. The presence of such proteins in yeast could indicate the existence of their homologues in mammalian cells and would greatly aid their identification. Expression of ClC-2 in yeast required optimization of the sequence context of the AUG translation initiation codon. After obtaining an efficient translation, we found that rat ClC-2 cannot directly substitute for yeast Gef1p. Functional substitution for Gef1p was, however, achieved in the presence of an increased level of intact or C-terminally truncated yeast Kha1 protein. Based on the deduced amino acid sequence, the Kha1 protein can be classified as a Na+/H+ transporter since it has a large N-terminal domain similar to the family of NHEs (Na+/H+ exchangers). This suggests that the Kha1p may take part in the regulation of intracellular cation homoeostasis and pH control. We have established that Kha1p is localized in the same cellular compartment as Gef1p and yeast-expressed ClC-2: the Golgi apparatus. We propose that Kha1p may aid ClC-2-dependent suppression of the Δgef1-assocciated growth defects by keeping the Golgi apparatus pH in a range suitable for ClC-2 activity. The approach employed in the present study may be of general applicability to the characterization of poorly understood proteins by their functional expression in yeast.

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