Functional Expression of House Fly (Musca domestica) Cytochrome P450 CYP6D1 in Yeast (Saccharomyces cerevisiae)

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.

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Expression, purification and direct eletrochemistry of cytochrome P450 6A1 from the house fly, Musca domestica

Protein Expression and Purification ◽

10.1016/j.pep.2009.12.008 ◽

2010 ◽

Vol 71 (1) ◽

pp. 74-78 ◽

Cited By ~ 15

Author(s):

Li Zhang ◽

Xuequn Liu ◽

Chuntai Wang ◽

Xinqiong Liu ◽

Gang Cheng ◽

...

Keyword(s):

Cytochrome P450 ◽

Musca Domestica ◽

House Fly

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Functional expression of recombinant N-methyl- D-aspartate receptors in the yeast Saccharomyces cerevisiae . Localization and pharmacological characterization

European Journal of Biochemistry ◽

10.1046/j.1432-1327.1998.2520391.x ◽

1998 ◽

Vol 252 (3) ◽

pp. 391-399 ◽

Cited By ~ 9

Author(s):

Ziyu Li ◽

Jurgen Becker ◽

Christian R. Noe

Keyword(s):

Saccharomyces Cerevisiae ◽

Functional Expression ◽

Yeast Saccharomyces Cerevisiae ◽

Pharmacological Characterization

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Biochemical changes in the cytochrome P450 monooxygenases of seven insecticide-resistant house fly (Musca domestica L.) strains

Pesticide Biochemistry and Physiology ◽

10.1016/0048-3575(90)90004-l ◽

1990 ◽

Vol 36 (2) ◽

pp. 127-134 ◽

Cited By ~ 20

Author(s):

Jeffrey G. Scott ◽

Susanna S.T. Lee ◽

Toshio Shono

Keyword(s):

Cytochrome P450 ◽

Musca Domestica ◽

House Fly ◽

Biochemical Changes ◽

Cytochrome P450 Monooxygenases ◽

P450 Monooxygenases

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

Molecular and Cellular Biology ◽

10.1128/mcb.7.6.2087-2096.1987 ◽

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.

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