Papers by Monique Bolotin-Fukuhara
Identification of a “Relic sequence” in <i>S. cerevisiae</i>
Functional comparison between <i>ScYAP1</i> and the <i>HpHAP4-B</i> genes
<p>Part A: Comparison of the new bZIP type motif identified in HpHap4-B with the YAP family... more <p>Part A: Comparison of the new bZIP type motif identified in HpHap4-B with the YAP family motif. Q234, Q239, A241 and F/Y242 are four basic regions (DNA-binding sites) characteristic residues of the YAP protein family which are rarely or never observed in other bZIP proteins <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0112263#pone.0112263-Fernandes1" target="_blank">[34]</a>. Part B: Heterologous complementation of the growth deficiency of <i>S. cerevisiae</i> Δ<i>yap1</i> in the presence of H<sub>2</sub>0<sub>2</sub> by the <i>HpHAP4-B</i> gene. The gene is expressed on a multicopy plasmid (pBFG1, see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0112263#s2" target="_blank"><i>Methods</i></a>) and the growth monitored on minimal medium (W0) containing 0.5 mM of H<sub>2</sub>0<sub>2</sub>. 1: <i>S. cerevisiae</i> Δ<i>yap1</i> strain (with or without H<sub>2</sub>0<sub>2</sub>). 2:Δ<i>yap1</i> strain with empty plasmid BFG1. 3: Δ<i>yap1</i> with BFG1 plasmid containing <i>HpHap4-B</i>. 4: Δ<i>yap1</i> with BFG1 plasmid containing <i>HpHap4-B</i> with deleted bZIP domain (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0112263#s2" target="_blank">Methods</a>). Tests 2, 3 and 4 were performed on medium containing H<sub>2</sub>0<sub>2</sub>. Strains were grown on minimal glucose medium supplemented with the necessary aminoacids at 28°C for 5 days.</p
A simple phylogenetic tree of the hemiascomycete yeasts
<p>Only a limited number of yeast species are presented which span the complete clade of He... more <p>Only a limited number of yeast species are presented which span the complete clade of Hemiascomycete yeasts from <i>S. cerevisiae</i> to <i>Y. lipolytica, S. pombe</i> being the outgroup. <i>H. polymorpha</i> is placed on the tree at the same position as <i>P. angusta</i> which is the anamorphic species. The position of the « Whole Genome Duplication » (WGD) is indicated. The tree is taken from <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0112263#pone.0112263-Souciet1" target="_blank">[48]</a>.</p
Growth of different <i>H. polymorpha</i> strains on media containing either antimycin A, SHAM or hydrogen peroxide
<p>A: <i>H. polymorpha Hp</i>Δ<i>hap4-A</i> and double knock-out st... more <p>A: <i>H. polymorpha Hp</i>Δ<i>hap4-A</i> and double knock-out strains, but not <i>Hp</i>Δ<i>hap4-B</i> deletion mutant, are sensitive to antimycin A on glucose. B: Growth on xylose plus SHAM or H<sub>2</sub>0<sub>2</sub>. 1: <i>Hp</i>NCYC495<i>leu1_1</i> strain. 2: <i>Hp</i>NCYC495<i>leu1_1</i> with pYT1 (empty plasmid, see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0112263#s2" target="_blank">Material and Methods</a>). 3: <i>Hp</i>Δ<i>hap4-A</i>. 4: <i>Hp</i>Δ<i>hap4-B</i>. 5: <i>Hp</i>Δ<i>hap4-A Hp</i>Δ<i>hap4-B</i> (double knock-out strain).</p
RESEARCH ARTICLE Efficient Mating-Type Switching in Candida glabrata Induces Cell Death
Candida glabrata is an apparently asexual haploid yeast that is phylogenetically closer to Saccha... more Candida glabrata is an apparently asexual haploid yeast that is phylogenetically closer to Saccharomyces cerevisiae than to Candida albicans. Its genome contains threeMAT-like cassettes,MAT, which encodes eitherMATa orMATalpha information in different strains, and the additional loci, HML and HMR. The genome also contains an HO gene homolog, but this yeast has never been shown to switch mating-types spontaneously, as S. cerevisiae does. We have recently sequenced the genomes of the five species that, together with C. glabrata, make up the Nakaseomyces clade. All containMAT-like cassettes and an HO gene homolog. In this work, we express the HO gene of all Nakaseomyces and of S. cerevi-siae inC. glabrata. All can induce mating-type switching, but, despite the larger phylogenetic distance, the most efficient endonuclease is the one from S. cerevisiae. Efficient mating-type switching in C. glabrata is accompanied by a high cell mortality, and sometimes results in conversion of the addit...
Yeast models of mitochondrial diseases due to mutations in nuclear genes
Mitochondrial diseases and the role oftheyeastmodels Teresa Rinaldi, Cristina Dallabona, Ileana F... more Mitochondrial diseases and the role oftheyeastmodels Teresa Rinaldi, Cristina Dallabona, Ileana Ferrero, Laura Frontali & Monique Bolotin-Fukuhara Department of Cell and Developmental Biology, Pasteur Institute-Cenci Bolognetti Foundation, Sapienza University of Rome, Rome, Italy; Department of Genetics, Biology of Microrganisms, Anthropology, Evolution, University of Parma, Parma, Italy; and Institut de Génétique et Microbiologie, CNRSUniversité Paris XI, Orsay, France
NAR Genomics and Bioinformatics, 2020
Candida glabrata is a cause of life-threatening invasive infections especially in elderly and imm... more Candida glabrata is a cause of life-threatening invasive infections especially in elderly and immunocompromised patients. Part of human digestive and urogenital microbiota, C. glabrata faces varying iron availability, low during infection or high in digestive and urogenital tracts. To maintain its homeostasis, C. glabrata must get enough iron for essential cellular processes and resist toxic iron excess. The response of this pathogen to both depletion and lethal excess of iron at 30°C have been described in the literature using different strains and iron sources. However, adaptation to iron variations at 37°C, the human body temperature and to gentle overload, is poorly known. In this study, we performed transcriptomic experiments at 30°C and 37°C with low and high but sub-lethal ferrous concentrations. We identified iron responsive genes and clarified the potential effect of temperature on iron homeostasis. Our exploration of the datasets was facilitated by the inference of functio...
Journal of Bacteriology, 1996
A gene for high-affinity glucose transport, HGT1, has been isolated from the lactose-assimilating... more A gene for high-affinity glucose transport, HGT1, has been isolated from the lactose-assimilating yeast Kluyveromyces lactis. Disruption strains showed much-reduced uptake of glucose at low concentrations and growth was particularly affected in low-glucose medium. The HGT1 nucleotide sequence implies that it encodes a typical transmembrane protein with 12 hydrophobic domains and with 26 to 31% amino acid identity with the Hxtp family of glucose transport elements in Saccharomyces cerevisiae. Expression is constitutive (in contrast to RAG1, the major gene for low-affinity glucose uptake in K. lactis) and is controlled by several genes also known to affect expression of RAG1. These include RAG5 (which codes for the single hexokinase of K. lactis), which is required for HGT1 transcription, and RAG4, which has a negative effect. The double mutant deltahgt1deltarag1 showed further reduced glucose uptake but still grew quite well on 2% glucose and was not completely impaired even on 0.1% ...
FEMS yeast research, 2016
FEMS Yeast Research, 2015
The decrease in carbon metabolic gene copy loss cannot be simply associated to a reduction of glu... more The decrease in carbon metabolic gene copy loss cannot be simply associated to a reduction of glucose consumption and can be counterbalanced by other beneficial genetic variations.
PLOS ONE, 2015
Candida glabrata is an apparently asexual haploid yeast that is phylogenetically closer to Saccha... more Candida glabrata is an apparently asexual haploid yeast that is phylogenetically closer to Saccharomyces cerevisiae than to Candida albicans. Its genome contains three MAT-like cassettes, MAT, which encodes either MATa or MATalpha information in different strains, and the additional loci, HML and HMR. The genome also contains an HO gene homolog, but this yeast has never been shown to switch mating-types spontaneously, as S. cerevisiae does. We have recently sequenced the genomes of the five species that, together with C. glabrata, make up the Nakaseomyces clade. All contain MAT-like cassettes and an HO gene homolog. In this work, we express the HO gene of all Nakaseomyces and of S. cerevisiae in C. glabrata. All can induce mating-type switching, but, despite the larger phylogenetic distance, the most efficient endonuclease is the one from S. cerevisiae. Efficient matingtype switching in C. glabrata is accompanied by a high cell mortality, and sometimes results in conversion of the additional cassette HML. Mortality probably results from the cutting of the HO recognition sites that are present, in HML and possibly HMR, contrary to what happens naturally in S. cerevisiae. This has implications in the life-cycle of C. glabrata, as we show that efficient MAT switching is lethal for most cells, induces chromosomal rearrangements in survivors, and that the endogenous HO is probably rarely active indeed.
PLoS ONE, 2009
The whole-genome duplication (WGD) may provide a basis for the emergence of the very characterist... more The whole-genome duplication (WGD) may provide a basis for the emergence of the very characteristic life style of Saccharomyces cerevisiae-its fermentation-oriented physiology and its capacity of growing in anaerobiosis. Indeed, we found an over-representation of oxygen-responding genes in the ohnologs of S. cerevisiae. Many of these duplicated genes are present as aerobic/hypoxic(anaerobic) pairs and form a specialized system responding to changing oxygen availability. HYP2/ANB1 and COX5A/COX5B are such gene pairs, and their unique orthologs in the 'non-WGD' Kluyveromyces lactis genome behaved like the aerobic versions of S. cerevisiae. ROX1 encodes a major oxygen-responding regulator in S. cerevisiae. The synteny, structural features and molecular function of putative KlROX1 were shown to be different from that of ROX1. The transition from the K. lactis-type ROX1 to the S. cerevisiae-type ROX1 could link up with the development of anaerobes in the yeast evolution. Bioinformatics and stochastic analyses of the Rox1p-binding site (YYYATTGTTCTC) in the upstream sequences of the S. cerevisiae Rox1p-mediated genes and of the K. lactis orthologs also indicated that K. lactis lacks the specific gene system responding to oxygen limiting environment, which is present in the 'post-WGD' genome of S. cerevisiae. These data suggested that the oxygen-responding system was born for the specialized physiology of S. cerevisiae.
PLoS ONE, 2014
The transcriptional regulator HAP4, induced by respiratory substrates, is involved in the balance... more The transcriptional regulator HAP4, induced by respiratory substrates, is involved in the balance between fermentation and respiration in S. cerevisiae. We identified putative orthologues of the Hap4 protein in all ascomycetes, based only on a conserved sixteen amino acid-long motif. In addition to this motif, some of these proteins contain a DNA-binding motif of the bZIP type, while being nonetheless globally highly divergent. The genome of the yeast Hansenula polymorpha contains two HAP4-like genes encoding the protein HpHap4-A which, like ScHap4, is devoid of a bZIP motif, and HpHap4-B which contains it. This species has been chosen for a detailed examination of their respective properties. Based mostly on global gene expression studies performed in the S. cerevisiae HAP4 disruption mutant (ScDhap4), we show here that HpHap4-A is functionally equivalent to ScHap4, whereas HpHap4-B is not. Moreover HpHAP4-B is able to complement the H 2 O 2 hypersensitivity of the ScYap1 deletant, YAP1 being, in S. cerevisiae, the main regulator of oxidative stress. Finally, a transcriptomic analysis performed in the
Antifolate screening using yeast expressing Plasmodium vivax dihydrofolate reductase and in vitro drug susceptibility assay for Plasmodium falciparum
Molecular and Biochemical Parasitology, 2007
The presence of homologous point mutations in the dhfr gene in Plasmodium vivax and Plasmodium fa... more The presence of homologous point mutations in the dhfr gene in Plasmodium vivax and Plasmodium falciparum is associated with resistance to antifolate drugs. The spread of antifolate resistance encouraged research for novel antifolate drugs active against both wild-type and dhfr-mutant strains of malaria parasites. Because P. vivax cannot be easily maintained in culture, we transformed a Saccharomyces cerevisiae DHFR-deleted mutant to express wild-type P. vivax dhfr gene and its mutant forms. Twenty-five dicyclic and tricyclic 2,4-diaminopyrimidine derivatives were screened. Six quinazoline compounds showed selective inhibition of yeast transformants expressing P. vivax dhfr genes. The 50% inhibitory concentration (IC(50)) of these six compounds was determined against field isolates of P. falciparum. Our results suggest that a close relationship between the yeast assay based on expression of P. vivax dhfr genes and the in vitro test using P. falciparum parasites in culture is a promising initial step for drug screening.
DNA sequence analysis of a 17 kb fragment of yeast chromosome XI physically localizes theMRB1 gene and reveals eight new open reading frames, including a homologue of the KIN1/KIN2 and SNF1 protein kinases
Yeast, 1993
We report in this paper the sequence of a part of chromosome XI of Saccharomyces cerevisiae. This... more We report in this paper the sequence of a part of chromosome XI of Saccharomyces cerevisiae. This 17 kbp nucleotide sequence represents the right half of cosmid pUKG151 and contains nine open reading frames, YKL453, 450, 449, 448, 445, 443, 442, 441 and the 5&#39; part of YKL440. YKL440 was previously identified as the MBR1 gene and plays a role in mitochondrial biogenesis. YKL443 is a homologue of the yeast serine-rich protein (SRP1), while YKL453 presents strong homologies with the KIN1/KIN2/SNF1 kinase family. It must be pointed out that the size of this gene is well above average for yeast.
FEMS Microbiology Letters, 2006
Sulpha drugs act as competitive inhibitors of p-amino benzoic acid, an intermediate in the de nov... more Sulpha drugs act as competitive inhibitors of p-amino benzoic acid, an intermediate in the de novo folate pathway. Dihydropteroate synthase condenses sulpha drugs into sulpha-dihydropteroate (sulpha-DHP), which competes with dihydrofolate, the dihydrofolate reductase (DHFR) substrate. This designates DHFR as a possible target of sulpha-DHP. We suggest here that Plasmodium vivax DHFR is indeed the in vivo target of sulpha drugs. The wild-type DHFR expressed in Saccharomyces cerevisiae leads to cell growth inhibition, while sensitivity to the drug is exacerbated in the mutants. Contrary to what is observed with sulphanilamide, methotrexate is less effective on P. vivax-DHFR mutants than on wild-type mutant.
Eukaryotic Cell, 2008
The HAP1 ( CYP1 ) gene product of Saccharomyces cerevisiae is known to regulate the transcription... more The HAP1 ( CYP1 ) gene product of Saccharomyces cerevisiae is known to regulate the transcription of many genes in response to oxygen availability. This response varies according to yeast species, probably reflecting the specific nature of their oxidative metabolism. It is suspected that a difference in the interaction of Hap1p with its target genes may explain some of the species-related variation in oxygen responses. As opposed to the fermentative S. cerevisiae , Kluyveromyces lactis is an aerobic yeast species which shows different oxygen responses. We examined the role of the HAP1 -equivalent gene (Kl HAP1 ) in K. lactis . KlHap1p showed a number of sequence features and some gene targets (such as Kl CYC1 ) in common with its S. cerevisiae counterpart, and Kl HAP1 was capable of complementing the hap1 mutation. However, the Kl HAP1 disruptant showed temperature-sensitive growth on glucose, especially at low glucose concentrations. At normal temperature, 28°C, the mutant grew wel...
MGG Molecular & General Genetics, 1987
Yeast strain 990 carries a mutation mapping to the olil locus of the mitochondrial genome, the ge... more Yeast strain 990 carries a mutation mapping to the olil locus of the mitochondrial genome, the gene encoding ATPase subunit 9. DNA sequence analysis indicated a substitution of valine for alanine at residue 22 of the protein. The strain failed to grow on nonfermentable carbon sources such as glycerol at low temperature (20 ° C). At 28 ° C the strain grew on nonfermentable carbon sources and was resistant to the antibiotic oligomycin. ATPase activity in mitochondria isolated from 990 was reduced relative to the wild-type strain from which it was derived, but the residual activity was oligomycin resistant. Subunit 9 (the DCCD-binding proteolipid) from the mutant strain exhibited reduced mobility in SDS-polyacrylamide gels relative to the wild-type proteolipid. Ten revertant strains of 990 were analyzed. All restored the ability to grow on glycerol at 20 ° C. Mitotic segregation data showed that eight of the ten revertants were attributable to mitochondrial genetic events and two were caused by nuclear events since they appeared to be recessive nuclear suppressors. These nuclear mutations retained partial resistance to oligomycin and did not alter the electrophoretic behavior of subunit 9 or any other ATPase subunit. When mitochondrial DNA from each of the revertant strains was hybridized with an oligonueleotide probe covering the olil mutation, seven of the mitochondrial revertants were found to be true reverrants and one a second mutation at the site of the original 990 mutation. The olil gene from this strain contained a substitution of glycine for valine at residue 22. The proteolipid isolated from this strain had increased electrophoretic mobility relative to the wild-type proteolipid.
Genetics, 1974
We have isolated 15 spontaneous mutants resistant to one or several antibiotics like chlorampheni... more We have isolated 15 spontaneous mutants resistant to one or several antibiotics like chloramphenicol, erythromycin and spiramycin. We have shown by several criteria that all of them result from mutations localized in the mitochondrial DNA. The mutations have been mapped by allelism tests and by two- and three-factor crosses involving various configurations of resistant and sensitive alleles associated in cis or in trans with the mitochondrial locus ω which governs the polarity of genetic recombination. A general mapping procedure based on results of heterosexual (ω+ × ω-) crosses and applicable to mutations localized in the polar segment is described and shown to be more resolving than that based on results of homosexual crosses. Mutations fall into three loci which are all linked and map in the following order: ω-RI-RII-RIII. The first locus is very tightly linked with ω while the second is less linked to the first. Mutations of similar resistance phenotype can belong to different ...
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Papers by Monique Bolotin-Fukuhara