christophe cullin's Publications List

Comparative studies of nontoxic and toxic amyloids interacting with membrane models at the air-water interface.

2011-09-05T16:53:07Z

Many in vitro studies have pointed out the interaction between amyloids and membranes, and their potential involvement in amyloid toxicity. In a previous study, we generated a yeast toxic mutant (M8) of the harmless model amyloid protein HET-s((218-289)). In this study, we compared the self-assembling process of the nontoxic wild-type (WT) and toxic (M8) protein at the air-water interface and in i...

Ha Phuong Ta, Karine Berthelot, Bénédicte Coulary-Salin, Bernard Desbat, Julie Géan, Laurent Servant, Christophe Cullin, Sophie Lecomte (2011) Langmuir 27: 8 4797-4807

InVivo and In Vitro Analyses of Toxic Mutants of HET-s: FTIR Antiparallel Signature Correlates with Amyloid Toxicity.

2011-09-05T16:53:07Z

The folding and interactions of amyloid proteins are at the heart of the debate as to how these proteins may or may not become toxic to their host. Although little is known about this issue, the structure seems to be clearly involved with effects on molecular events. To understand how an amyloid may be toxic, we previously generated a yeast toxic amyloid (mutant 8) from the nontoxic HET-s((218-289...

Karine Berthelot, Ha Phuong Ta, Julie Géan, Sophie Lecomte, Christophe Cullin (2011) J Mol Biol 412: 1 137-152

The toxicity of an "artificial" amyloid is related to how it interacts with membranes.

2011-09-05T16:53:07Z

Despite intensive research into how amyloid structures can impair cellular viability, the molecular nature of these toxic species and the cellular mechanisms involved are not clearly defined and may differ from one disease to another. We systematically analyzed, in Saccharomyces cerevisiae, genes that increase the toxicity of an amyloid (M8), previously selected in yeast on the sole basis of its c...

Julien Couthouis, Christelle Marchal, Fabien D'Angelo, Karine Berthelot, Christophe Cullin (2010) Prion 4: 4 283-291

A yeast toxic mutant of HET-s((218-289)) prion displays alternative intermediates of amyloidogenesis.

2011-09-05T16:53:07Z

Amyloids are thought to be involved in various types of neurodegenerative disorders. Several kinds of intermediates, differing in morphology, size, and toxicity, have been identified in the multistep amyloidogenesis process. However, the mechanisms explaining amyloid toxicity remain unclear. We previously generated a toxic mutant of the nontoxic HET-s((218-289)) amyloid in yeast. Here we report th...

Karine Berthelot, Sophie Lecomte, Julie Géan, Françoise Immel, Christophe Cullin (2010) Biophys J 99: 4 1239-1246

The polyphenol piceid destabilizes preformed amyloid fibrils and oligomers in vitro: hypothesis on possible molecular mechanisms.

2011-09-05T16:53:07Z

Alzheimer's disease (AD) is characterized by deposits of amyloid in various tissues. The neuronal cytotoxicity of Abeta peptides is attributed not only to various mechanisms but also to amyloid fibrils and soluble oligomeric intermediates. Consequently, finding molecules to prevent or reverse the oligomerization and fibrillization of Abeta could be of therapeutic value in the treatment of AD. We s...

Céline Rivière, Jean-Claude Delaunay, Françoise Immel, Christophe Cullin, Jean-Pierre Monti (2009) Neurochem Res 34: 6 1120-1128

The cellular concentration of the yeast Ure2p prion protein affects its propagation as a prion.

2011-09-05T16:53:07Z

The [URE3] yeast prion is a self-propagating inactive form of the Ure2p protein. We show here that Ure2p from the species Saccharomyces paradoxus (Ure2p(Sp)) can be efficiently converted into a prion form and propagate [URE3] when expressed in Saccharomyces cerevisiae at physiological level. We found however that Ure2p(Sp) overexpression prevents efficient prion propagation. We have compared the a...

Myriam Crapeau, Christelle Marchal, Christophe Cullin, Laurent Maillet (2009) Mol Biol Cell 20: 8 2286-2296

Driving amyloid toxicity in a yeast model by structural changes: a molecular approach.

2011-09-05T16:53:07Z

The amyloid aggregation pathway is a multistep process, and many in vitro studies have highlighted the role of particular intermediates in the cellular toxicity of various amyloid diseases. In a previous study, we generated a yeast toxic mutant (M8) of the harmless model amyloid protein Het-s(218-289). In this study, we compared the aggregation characteristics of the wild-type (WT) and the toxic m...

Karine Berthelot, Françoise Immel, Julie Géan, Sophie Lecomte, Reiko Oda, Brice Kauffmann, Christophe Cullin (2009) FASEB J 23: 7 2254-2263

Screening for toxic amyloid in yeast exemplifies the role of alternative pathway responsible for cytotoxicity.

2011-09-05T16:53:07Z

The relationship between amyloid and toxic species is a central problem since the discovery of amyloid structures in different diseases. Despite intensive efforts in the field, the deleterious species remains unknown at the molecular level. This may reflect the lack of any structure-toxicity study based on a genetic approach. Here we show that a structure-toxicity study without any biochemical pre...

Julien Couthouis, Karine Rébora, Françoise Immel, Karine Berthelot, Michel Castroviejo, Christophe Cullin (2009) PLoS One 4: 3

Nitrogen catabolic repression controls the release of volatile thiols by Saccharomyces cerevisiae during wine fermentation.

2011-09-05T16:53:07Z

Volatile thiols such as 4-methyl-4-sulfanylpentan-2-one (4MSP) and 3-sulfanylhexan-1-ol (3SH) are aromatic molecules having an important organoleptic impact on white wines. These components are produced from inodorous nonvolatile cysteinylated precursors by Saccharomyces cerevisiae metabolic activity during alcoholic fermentation. Here we provide a new insight into the genetic determinism of the p...

Cécile Thibon, Philippe Marullo, Olivier Claisse, Christophe Cullin, Denis Dubourdieu, Takatoshi Tominaga (2008) FEMS Yeast Res 8: 7 1076-1086

In vitro analysis of SpUre2p, a prion-related protein, exemplifies the relationship between amyloid and prion.

2011-09-05T16:53:07Z

The yeast Saccharomyces cerevisiae contains in its proteome at least three prion proteins. These proteins (Ure2p, Sup35p, and Rnq1p) share a set of remarkable properties. In vivo, they form aggregates that self-perpetuate their aggregation. This aggregation is controlled by Hsp104, which plays a major role in the growth and severing of these prions. In vitro, these prion proteins form amyloid fibr...

Francoise Immel, Yi Jiang, Yi-Qian Wang, Christelle Marchal, Laurent Maillet, Sarah Perrett, Christophe Cullin (2007) J Biol Chem 282: 11 7912-7920

A novel in vitro filter trap assay identifies tannic acid as an amyloid aggregation inducer for HET-s.

2011-09-05T16:53:07Z

In this work we present an easy and low cost in vitro filter trap assay to quickly identify direct actors on amyloid prion aggregation. We chose the recombinant purified prion protein HET-s from Podospora anserina as a reference. HET-s was labelled with a fluorophore prior to aggregation assays in a 96 well micro-array system. Aggregation assays were carried out in presence of a number of chemical...

Mona Boyé-Harnasch, Christophe Cullin (2006) J Biotechnol 125: 2 222-230

The [URE3] prion is not conserved among Saccharomyces species.

2011-09-05T16:53:07Z

The [URE3] prion of Saccharomyces cerevisiae is a self-propagating inactive form of the nitrogen catabolism regulator Ure2p. To determine whether the [URE3] prion is conserved in S. cerevisiae-related yeast species, we have developed genetic tools allowing the detection of [URE3] in Saccharomyces paradoxus and Saccharomyces uvarum. We found that [URE3] is conserved in S. uvarum. In contrast, [URE3...

Nicolas Talarek, Laurent Maillet, Christophe Cullin, Michel Aigle (2005) Genetics 171: 1 23-34

The [URE3] yeast prion results from protein aggregates that differ from amyloid filaments formed in vitro.

2011-09-05T16:53:07Z

The [URE3] yeast prion is a self-propagating inactive form of the Ure2 protein. Ure2p is composed of two domains, residues 1-93, the prion-forming domain, and the remaining C-terminal part of the protein, which forms the functional domain involved in nitrogen catabolite repression. In vitro, Ure2p forms amyloid filaments that have been proposed to be the aggregated prion form found in vivo. Here w...

Leslie Ripaud, Laurent Maillet, Françoise Immel-Torterotot, Fabien Durand, Christophe Cullin (2004) J Biol Chem 279: 49 50962-50968

Developing methods and strains for genetic studies in the Saccharomyces bayanus var. uvarum species.

2011-09-05T16:53:07Z

For years, Saccharomyces cerevisiae has been used as a model organism to gain insight into complex biological processes. The study of closely related yeast species may be critical for understanding the molecular mechanism of evolution. Among those species, S. bayanus var. uvarum could be particularly pertinent because of the availability of its genome sequence. However, to date, in that species ge...

Nicolas Talarek, Edward J Louis, Christophe Cullin, Michel Aigle (2004) Yeast 21: 14 1195-1203

Internal initiation drives the synthesis of Ure2 protein lacking the prion domain and affects [URE3] propagation in yeast cells.

2011-09-05T16:53:07Z

The [URE3] phenotype in Saccharomyces cerevisiae is caused by the inactive, altered (prion) form of the Ure2 protein (Ure2p), a regulator of nitrogen catabolism. Ure2p has two functional domains: an N-terminal domain necessary and sufficient for prion propagation and a C-terminal domain responsible for nitrogen regulation. We show here that the mRNA encoding Ure2p possesses an IRES (internal ribos...

Anton A Komar, Thierry Lesnik, Christophe Cullin, William C Merrick, Hans Trachsel, Michael Altmann (2003) EMBO J 22: 5 1199-1209

Isolation of drugs active against mammalian prions using a yeast-based screening assay.

2011-09-05T16:53:07Z

We have developed a rapid, yeast-based, two-step assay to screen for antiprion drugs. The method allowed us to identify several compounds effective against budding yeast prions responsible for the [PSI+] and [URE3] phenotypes. These inhibitors include the kastellpaolitines, a new class of compounds, and two previously known molecules, phenanthridine and 6-aminophenanthridine. Two potent promoters ...

Stéphane Bach, Nicolas Talarek, Thibault Andrieu, Jean-Michel Vierfond, Yvette Mettey, Hervé Galons, Dominique Dormont, Laurent Meijer, Christophe Cullin, Marc Blondel (2003) Nat Biotechnol 21: 9 1075-1081

The mechanisms of [URE3] prion elimination demonstrate that large aggregates of Ure2p are dead-end products.

2011-09-05T16:53:07Z

The yeast prion [URE3] is a self-propagating inactive form (the propagon) of the Ure2 protein. Ure2p is composed of two domains: residues 1-93--the prion-forming domain (PFD)--and the remaining C-terminal part of the protein, which forms the functional domain involved in nitrogen catabolite repression. Guanidine hydrochloride, and the overproduction of Ure2p 1-65 or Ure2-GFP have been shown to ind...

Leslie Ripaud, Laurent Maillet, Christophe Cullin (2003) EMBO J 22: 19 5251-5259

Conservation of the prion properties of Ure2p through evolution.

2011-09-05T16:53:07Z

The yeast inheritable [URE3] element corresponds to a prion form of the nitrogen catabolism regulator Ure2p. We have isolated several orthologous URE2 genes in different yeast species: Saccharomyces paradoxus, S. uvarum, Kluyveromyces lactis, Candida albicans, and Schizosaccharomyces pombe. We show here by in silico analysis that the GST-like functional domain and the prion domain of the Ure2 prot...

Agnès Baudin-Baillieu, Eric Fernandez-Bellot, Fabienne Reine, Eric Coissac, Christophe Cullin (2003) Mol Biol Cell 14: 8 3449-3458

The [URE3] phenotype: evidence for a soluble prion in yeast.

2011-09-05T16:53:07Z

The aggregation of the two yeast proteins Sup35p and Ure2p is widely accepted as a model for explaining the prion propagation of the phenotypes [PSI+] and [URE3], respectively. Here, we demonstrate that the propagation of [URE3] cannot simply be the consequence of generating large aggregates of Ure2p, because such aggregation can be found in some conditions that are not related to the prion state ...

Eric Fernandez-Bellot, Elisabeth Guillemet, Frederique Ness, Agnes Baudin-Baillieu, Leslie Ripaud, Mick Tuite, Christophe Cullin (2002) EMBO Rep 3: 1 76-81

The protein-only theory and the yeast Saccharomyces cerevisiae: the prions and the propagons.

2011-09-05T16:53:07Z

The yeast prions represent a very attractive and tractable model for investigating the prion world. The more extensively studied yeast prion [PSI] leads to a propagation model that links auto-aggregation in amyloid formation and inactivation of the cellular function of the yeast 'prion protein' Sup35p. The other prion model, [URE3], appears to be similar in some genetic and biochemical properties....

E Fernandez-Bellot, C Cullin (2001) Cell Mol Life Sci 58: 12-13 1857-1878

Translational errors as an early event in prion conversion.

2011-09-05T16:53:07Z

A prion is an infectious, altered form of a cellular protein which can self-propagate and affect normal phenotype. Prion conversion has been observed for mammalian and yeast proteins but molecular mechanisms that trigger this process remain unclear. Up to now, only post-translational models have been explored. In this work, we tested the hypothesis that co-translational events may be implicated in...

I Hatin, L Bidou, C Cullin, J P Rousset (2001) Cell Mol Biol (Noisy-le-grand) 47 Online Pub: OL23-OL28

Stability, folding, dimerization, and assembly properties of the yeast prion Ure2p.

2011-09-05T16:53:07Z

The [URE3] factor of Saccharomyces cerevisiae propagates by a prion-like mechanism and corresponds to the loss of the function of the cellular protein Ure2. The molecular basis of the propagation of this phenotype is unknown. We recently expressed Ure2p in Escherichia coli and demonstrated that the N-terminal region of the protein is flexible and unstructured, while its C-terminal region is compac...

C Thual, L Bousset, A A Komar, S Walter, J Buchner, C Cullin, R Melki (2001) Biochemistry 40: 6 1764-1773

The yeast prion [URE3] can be greatly induced by a functional mutated URE2 allele.

2011-09-05T16:53:07Z

The non-Mendelian element [URE3] of yeast is considered to be a prion form of the Ure2 protein. The [URE3] phenotype occurs at a frequency of 10(-5) in haploid yeast strains, is reversible, and its frequency is increased by overexpressing the URE2 gene. We created a new mutant of the Ure2 protein, called H2p, which results in a 1000-fold increase in the rate of [URE3] occurrence. To date, only the...

E Fernandez-Bellot, E Guillemet, C Cullin (2000) EMBO J 19: 13 3215-3222

Structural characterization of Saccharomyces cerevisiae prion-like protein Ure2.

2011-09-05T16:53:07Z

Sacchromyces cerevisiae prion-like protein Ure2 was expressed in Escherichia coli and was purified to homogeneity. We show here that Ure2p is a soluble protein that can assemble into fibers that are similar to the fibers observed in the case of PrP in its scrapie prion filaments form or that form on Sup35 self-assembly. Ure2p self-assembly is a cooperative process where one can distinguish a lag p...

C Thual, A A Komar, L Bousset, E Fernandez-Bellot, C Cullin, R Melki (1999) J Biol Chem 274: 19 13666-13674

The [URE3] yeast prion: from genetics to biochemistry.

2011-09-05T16:53:07Z

[URE3] is a non-Mendelian genetic element of the yeast Saccharomyces cerevisiae, an altered prion form of Ure2 protein. We show that recombinant Ure2p is a soluble protein that can assemble in vitro into dimers, tetramers, and octamers or form insoluble fibrils observed for PrP in its filamentous form or for Sup35p upon self-assembling, suggesting a similar mechanism for all prions. Computational,...

A A Komar, R Melki, C Cullin (1999) Biochemistry (Mosc) 64: 12 1401-1407

Characterization of the interaction domains of Ure2p, a prion-like protein of yeast.

2011-09-05T16:53:07Z

In the yeast Saccharomyces cerevisiae, the non-Mendelian inherited genetic element [URE3] behaves as a prion. A hypothesis has been put forward which states that [URE3] arises spontaneously from its cellular isoform Ure2p (the product of the URE2 gene), and propagates through interactions of the N-terminal domain of the protein, thus leading to its aggregation and loss of function. In the present ...

E Fernandez-Bellot, E Guillemet, A Baudin-Baillieu, S Gaumer, A A Komar, C Cullin (1999) Biochem J 338 ( Pt 2): 403-407

Enhanced expression of the yeast Ure2 protein in Escherichia coli: the effect of synonymous codon substitutions at a selected place in the gene.

2011-09-05T16:53:07Z

The expression of the yeast Ure2 protein and its two N- and C-terminal HA-(YPYPVDYA) epitope and His-tag fusions has been enhanced in E. coli by selected silent mutagenesis of the URE2 gene. The two Arg-AGA codons at positions 253 and 254 of the URE2 gene coding sequence were exchanged by CGT codons accordingly. This has allowed an increased yield (up to 100-fold) of the full-length protein synthe...

A A Komar, E Guillemet, C Reiss, C Cullin (1998) Biol Chem 379: 10 1295-1300

Construction of a yeast strain deleted for the TRP1 promoter and coding region that enhances the efficiency of the polymerase chain reaction-disruption method.

2011-09-05T16:53:07Z

The sequence of the genome of Saccharomyces cerevisiae was recently determined. As well as all the informations concerning the structure of the chromosomes the scientific community had to deal with the discovery of dozens of new open reading frames (ORFs) of unknown function. The study of these ORFs requires the development of simple procedures that can be used on a large scale. In the framework o...

A Baudin-Baillieu, E Guillemet, C Cullin, F Lacroute (1997) Yeast 13: 4 353-356

Functional analysis of Rrp7p, an essential yeast protein involved in pre-rRNA processing and ribosome assembly.

2011-09-05T16:53:07Z

During the functional analysis of open reading frames (ORFs) identified during the sequencing of chromosome III of Saccharomyces cerevisiae, the previously uncharacterized ORF YCL031C (now designated RRP7) was deleted. RRP7 is essential for cell viability, and a conditional null allele was therefore constructed, by placing its expression under the control of a regulated GAL promoter. Genetic deple...

A Baudin-Baillieu, D Tollervey, C Cullin, F Lacroute (1997) Mol Cell Biol 17: 9 5023-5032

Differential resistance to proteinase K digestion of the yeast prion-like (Ure2p) protein synthesized in vitro in wheat germ extract and rabbit reticulocyte lysate cell-free translation systems.

2011-09-05T16:53:07Z

The Ure2p yeast prion-like protein was translated in vitro in the presence of labeled [35S]methionine in either rabbit reticulocyte lysate (RRL) or wheat germ extract (WGE) cell-free systems. When subjected to proteinase K digestion, the Ure2p protein synthesized in WGE was proteolysed much more slowly compared to that synthesized in RRL; this displays fragments of about 31-34 kDa, persisting over...

A A Komar, T Lesnik, C Cullin, E Guillemet, R Ehrlich, C Reiss (1997) FEBS Lett 415: 1 6-10

Functional analysis of YCL09C: evidence for a role as the regulatory subunit of acetolactate synthase.

2011-09-05T16:53:07Z

We have analysed the function of the open reading frame (ORF) YCL09C. The deletion of this ORF from chromosome III does not affect the physiology of the corresponding yeast strain enough to give a distinct phenotype. Nevertheless a computational analysis reveals high homology between this ORF and the enterobacterial genes encoding the regulatory subunit of acetolactate synthase. We have therefore ...

C Cullin, A Baudin-Baillieu, E Guillemet, O Ozier-Kalogeropoulos (1996) Yeast 12: 15 1511-1518

Multipurpose vectors designed for the fast generation of N- or C-terminal epitope-tagged proteins.

2011-09-05T16:53:07Z

In this paper are described a set of new high-copy-number yeast vectors, which are specially designed for the conditional expression of epitope-tagged proteins in vivo. One of the major advantages of these plasmids is that they allow polymerase chain reaction-amplified open reading frames to be automatically fused in frame with the epitope-coding sequence, avoiding longer procedures such as site-d...

C Cullin, L Minvielle-Sebastia (1994) Yeast 10: 1 105-112

Cellular localization of RNA14p and RNA15p, two yeast proteins involved in mRNA stability.

2011-09-05T16:53:07Z

RNA14 and RNA15 were originally identified by temperature-sensitive mutations that cause a rapid decrease in poly(A)-tail length and overall mRNA levels at the restrictive temperature. We have raised antibodies to the RNA14 and RNA15 proteins, and used subcellular fractionation and immunofluorescence to localize these proteins within the yeast cell. RNA14p is a 73 kDa protein found in both the nuc...

N Bonneaud, L Minvielle-Sebastia, C Cullin, F Lacroute (1994) J Cell Sci 107 ( Pt 4): 913-921

Cloning and characterization of a yeast cytochrome b5-encoding gene which suppresses ketoconazole hypersensitivity in a NADPH-P-450 reductase-deficient strain.

2011-09-05T16:53:07Z

Cytochrome P-450 (Cyp) 51 or lanosterol-C14-demethylase is the main target for antifungal compounds of the triazole family like ketoconazole (Kz). Disruption of the associated NADPH-P-450 reductase-encoding gene (YRED) is not lethal, but decreases by about 20-fold the Kz resistance (KzR) of wild-type (wt) Saccharomyces cerevisiae. Transformation of a YRED-disrupted strain by a yeast genomic librar...

G Truan, J C Epinat, C Rougeulle, C Cullin, D Pompon (1994) Gene 142: 1 123-127

The sequence of 29.7 kb from the right arm of chromosome II reveals 13 complete open reading frames, of which ten correspond to new genes.

2011-09-05T16:53:07Z

We have determined the complete nucleotide sequence of a 29.7 kb segment from the right arm of chromosome II carried by the cosmid alpha 61. The sequence encodes the 3' region of the IRA1 gene and 13 complete open reading frames, of which ten correspond to new genes and three (CIF1, ATPsv and CKS1) have been sequenced previously. The density of protein coding sequences is particularly high and cor...

A M Bécam, C Cullin, E Grzybowska, F Lacroute, F Nasr, O Ozier-Kalogeropoulos, A Palucha, P P Slonimski, M Zagulski, C J Herbert (1994) Yeast 10 Suppl A: S1-11

Enhanced in vivo monooxygenase activities of mammalian P450s in engineered yeast cells producing high levels of NADPH-P450 reductase and human cytochrome b5.

2011-09-05T16:53:07Z

We have engineered yeast genomic DNA to construct a set of strains producing various relative amounts of yeast NADPH-P450 reductase (Yred) and human cytochrome b5 (Hb5). Expression of cDNAs encoding human P450 1A1, 1A2, 3A4, 19A and mouse P450 1A1 in the different oxido-reduction backgrounds thus constituted were achieved after strain transformation by plasmid-based P450-encoding expression casset...

G Truan, C Cullin, P Reisdorf, P Urban, D Pompon (1993) Gene 125: 1 49-55

Recombinant yeast in drug metabolism.

2011-09-05T16:53:07Z

The usefulness of cDNA-directed expression of human hepatic P450s in yeast for the in vitro study of drug metabolism is emphasized. The major advantages of yeast expression are: (i) relatively high yields of heterologous P450 (approximately 5-10 nmol/l of culture medium) can be obtained; (ii) the expressed P450s are directly active in yeast microsomes, allowing the determination of specific cataly...

J P Renaud, M A Peyronneau, P Urban, G Truan, C Cullin, D Pompon, P Beaune, D Mansuy (1993) Toxicology 82: 1-3 39-52

A simple and efficient method for direct gene deletion in Saccharomyces cerevisiae.

2011-09-05T16:53:07Z

A Baudin, O Ozier-Kalogeropoulos, A Denouel, F Lacroute, C Cullin (1993) Nucleic Acids Res 21: 14 3329-3330

Expression in yeast of three allelic cDNAs coding for human liver P-450 3A4. Different stabilities, binding properties and catalytic activities of the yeast-produced enzymes.

2011-09-05T16:53:07Z

Three natural allelic cDNAs coding for P-450 3A4, the major form in human liver, namely NF25, NF10 and hPCN1, have been expressed in Saccharomyces cerevisiae. NF25 and hPCN1 were functionally expressed in yeast microsomes, yielding proteins with an absorption maximum at 448 nm in the CO-reduced difference spectrum. Some catalytic activities and substrate binding properties of P-450 NF25 and P-450 ...

M A Peyronneau, J P Renaud, M Jaouen, P Urban, C Cullin, D Pompon, D Mansuy (1993) Eur J Biochem 218: 2 355-361

Two distinct sequences control the targeting and anchoring of the mouse P450 1A1 into the yeast endoplasmic reticulum membrane.

2011-09-05T16:53:07Z

We previously expressed mouse P450 1A1 in the yeast S. cerevisiae. In the present study, I describe experiments in which several deletions in the 5' end of the corresponding cDNA were created. The truncated forms were then expressed in yeast cells. Studies of microsomes obtained from transformed yeast show that the signal-sequence is not required in vivo for the integration of mouse P450 1A1 into ...

C Cullin (1992) Biochem Biophys Res Commun 184: 3 1490-1495

Maximizing the expression of mammalian cytochrome P-450 monooxygenase activities in yeast cells.

2011-09-05T16:53:07Z

Cytochrome P-450s constitute a superfamily of mono-oxygenases which require the association with specific redox enzymes bound to the endoplasmic reticulum membrane for their activity. Conditions for the functional expression of these mammalian enzymes in yeast cells and the respective merits and limitations of currently used P-450 expression systems, are considered. The dependence of the mouse P-4...

P Urban, C Cullin, D Pompon (1990) Biochimie 72: 6-7 463-472

Expression of human liver cytochrome P450 IIIA4 in yeast. A functional model for the hepatic enzyme.

2011-09-05T16:53:07Z

Cytochrome P-450 (P450) NF, a member of the P450 IIIA subfamily, is the major contributor to the oxidation of the calcium-channel blocker nifedipine in human liver microsomes. A cDNA clone designated NF25 encoding for human P450 NF was isolated from a bacteriophage lambda gt11 expression library [Beaune, P. H., Umbenhauer, D. R., Bork, R. W., Lloyd, R. S. & Guengerich, F. P. (1986) Proc. Natl Acad...

J P Renaud, C Cullin, D Pompon, P Beaune, D Mansuy (1990) Eur J Biochem 194: 3 889-896

Synthesis of functional mouse cytochromes P-450 P1 and chimeric P-450 P3-1 in the yeast Saccharomyces cerevisiae.

2011-09-05T16:53:07Z

Mouse liver cytochrome P-450 P1 was produced in the yeast Saccharomyces cerevisiae transformed by various expression vectors. The relative efficiency of the phosphoglycerate kinase and GAL10-CYC1 promoters to direct the P-450 P1 mRNA synthesis was determined. The level of protein synthesis was found to be dependent on the amount of the 5'-noncoding sequence of the original cDNA removed during the ...

C Cullin, D Pompon (1988) Gene 65: 2 203-217