Proteasome- and SCF-dependent degradation of yeast adenine deaminase upon transition from proliferation to quiescence requires a new F-box protein named Saf1p.

In response to nutrient limitation, Saccharomyces cerevisiae cells enter into a nonproliferating state termed quiescence. This transition is associated with profound changes in gene expression patterns. The adenine deaminase encoding gene AAH1 is among the most precociously and tightly down-regulated gene upon entry into quiescence. We show that AAH1 down regulation is not specifically due to glucose exhaustion but is a more general response to nutrient limitation. We also found that Aah1p level is tightly correlated to RAS activity indicating thus an important role for the proteine kinase A pathway in this regulation process. We have isolated three deletion mutants, srb10, srb11 and saf1 (ybr280c) affecting AAH1 expression during post-diauxic growth and in early stationary phase. We show that the Srb10p cyclin-dependent kinase and its cyclin, Srb11p, regulate AAH1 expression at the transcriptional level. By contrast, Saf1p, a previously uncharacterized F-box protein, acts at a post-transcriptional level by promoting degradation of Aah1p. This post-transcriptional regulation is abolished by mutations affecting the proteasome or constant subunits of the SCF (Skp1-Cullin-F-box) complex. We propose that Saf1p targets Aah1p for proteasomedependent degradation upon entry into quiescence. This work provides the first direct evidence for active degradation of proteins in quiescent yeast cells

Multiple chemo-genetic interactions between a toxic metabolite and the ubiquitin pathway in yeast

International audienc

The Dct−/− Mouse Model to Unravel Retinogenesis Misregulation in Patients with Albinism

We have recently identified encoding dopachrome tautomerase (DCT) as the eighth gene for oculocutaneous albinism (OCA). Patients with loss of function of suffer from eye hypopigmentation and retinal dystrophy. Here we investigate the eye phenotype in mice. We show that their retinal pigmented epithelium (RPE) is severely hypopigmented from early stages, contrasting with the darker melanocytic tissues. Multimodal imaging reveals specific RPE cellular defects. Melanosomes are fewer with correct subcellular localization but disrupted melanization. RPE cell size is globally increased and heterogeneous. P-cadherin labeling of newborn RPE reveals a defect in adherens junctions similar to what has been described in tyrosinase-deficient embryos. The first intermediate of melanin biosynthesis, dihydroxyphenylalanine (L-Dopa), which is thought to control retinogenesis, is detected in substantial yet significantly reduced amounts in postnatal mouse eyecups. L-Dopa synthesis in the RPE alone remains to be evaluated during the critical period of retinogenesis. The mouse should prove useful in understanding the molecular regulation of retinal development and aging of the hypopigmented eye. This may guide therapeutic strategies to prevent vision deficits in patients with albinism.Approches de génétique moléculaire et fonctionnelle pour déchiffrer les mécanismes physiopathologiques de l'albinisme oculocutané

Disruption of Nucleotide Homeostasis by the Antiproliferative Drug 5-Aminoimidazole-4-carboxamide-1-β-d-ribofuranoside Monophosphate (AICAR)

International audienceBackground: AICAR is a potent anti-proliferative compound, but the basis of its cytotoxicity is poorly understood. Results: AICAR affects NTP homeostasis in a carbon source-dependent way, in both yeast and human cells. Conclusion: AICAR balance with nucleotides triphosphate is critical for its in vivo effects. Significance: AICAR is significantly more cytotoxic on glucose and thus potentially targets cells prone to Warburg effect

The SPX domain of the yeast low-affinity phosphate transporter Pho90 regulates transport activity

Yeast has two phosphate-uptake systems that complement each other: the high-affinity transporters (Pho84 and Pho89) are active under phosphate starvation, whereas Pho87 and Pho90 are low-affinity transporters that function when phosphate is abundant. Here, we report new regulatory functions of the amino-terminal SPX domain of Pho87 and Pho90. By studying truncated versions of Pho87 and Pho90, we show that the SPX domain limits the phosphate-uptake velocity, suppresses phosphate efflux and affects the regulation of the phosphate signal transduction pathway. Furthermore, split-ubiquitin assays and co-immunoprecipitation suggest that the SPX domain of both Pho90 and Pho87 interacts physically with the regulatory protein Spl2. This work suggests that the SPX domain inhibits low-affinity phosphate transport through a physical interaction with Spl2

Metabolic status rather than cell cycle signals control quiescence entry and exit

The use of new candidate markers for yeast quiescence reveals that quiescence entry and exit primarily rely on cellular metabolic status and can be uncoupled from the cell cycle

The Necrotic Signal Induced by Mycophenolic Acid Overcomes Apoptosis-Resistance in Tumor Cells

The amount of inosine monophosphate dehydrogenase (IMPDH), a pivotal enzyme for the biosynthesis of the guanosine tri-phosphate (GTP), is frequently increased in tumor cells. The anti-viral agent ribavirin and the immunosuppressant mycophenolic acid (MPA) are potent inhibitors of IMPDH. We recently showed that IMPDH inhibition led to a necrotic signal requiring the activation of Cdc42.Herein, we strengthened the essential role played by this small GTPase in the necrotic signal by silencing Cdc42 and by the ectopic expression of a constitutive active mutant of Cdc42. Since resistance to apoptosis is an essential step for the tumorigenesis process, we next examined the effect of the MPA–mediated necrotic signal on different tumor cells demonstrating various mechanisms of resistance to apoptosis (Bcl2-, HSP70-, Lyn-, BCR-ABL–overexpressing cells). All tested cells remained sensitive to MPA–mediated necrotic signal. Furthermore, inhibition of IMPDH activity in Chronic Lymphocytic Leukemia cells was significantly more efficient at eliminating malignant cells than apoptotic inducers.These findings indicate that necrosis and apoptosis are split signals that share few if any common hub of signaling. In addition, the necrotic signaling pathway induced by depletion of the cellular amount of GTP/GDP would be of great interest to eliminate apoptotic-resistant tumor cells

Effects of eight neuropsychiatric copy number variants on human brain structure

Many copy number variants (CNVs) confer risk for the same range of neurodevelopmental symptoms and psychiatric conditions including autism and schizophrenia. Yet, to date neuroimaging studies have typically been carried out one mutation at a time, showing that CNVs have large effects on brain anatomy. Here, we aimed to characterize and quantify the distinct brain morphometry effects and latent dimensions across 8 neuropsychiatric CNVs. We analyzed T1-weighted MRI data from clinically and non-clinically ascertained CNV carriers (deletion/duplication) at the 1q21.1 (n = 39/28), 16p11.2 (n = 87/78), 22q11.2 (n = 75/30), and 15q11.2 (n = 72/76) loci as well as 1296 non-carriers (controls). Case-control contrasts of all examined genomic loci demonstrated effects on brain anatomy, with deletions and duplications showing mirror effects at the global and regional levels. Although CNVs mainly showed distinct brain patterns, principal component analysis (PCA) loaded subsets of CNVs on two latent brain dimensions, which explained 32 and 29% of the variance of the 8 Cohen’s d maps. The cingulate gyrus, insula, supplementary motor cortex, and cerebellum were identified by PCA and multi-view pattern learning as top regions contributing to latent dimension shared across subsets of CNVs. The large proportion of distinct CNV effects on brain morphology may explain the small neuroimaging effect sizes reported in polygenic psychiatric conditions. Nevertheless, latent gene brain morphology dimensions will help subgroup the rapidly expanding landscape of neuropsychiatric variants and dissect the heterogeneity of idiopathic conditions

Mise en évidence d'une nouvelle voie impliquée dans l'homéostasie de la taille cellulaire chez S. cerevisiae

L homéostasie de la taille des cellules implique l existence de mécanismes capables de coordonner la croissance (l augmentation du volume) avec la prolifération (l augmentation du nombre de cellules). Il est clairement établi que la taille des cellules est affectée par la disponibilité en nutriment du milieu de culture et par la ploïdie, mais les mécanismes sous-jacents demeurent inconnus. Une étude à l échelle du génome réalisée chez la levure par l équipe de M. Tyers a révélée que l inactivation d environ 400 gènes conduisait à un volume cellulaire moyen significativement différent de celui de la souche sauvage isogénique. Le contrôle de la taille des cellules est ainsi une situation intéressante dans laquelle de nombreux loci contribuent à un caractère quantitatif complexe. La plupart de ces loci demeurant orphelin de voie de signalisation distincte, leur influence respective reste à élucider. Nous avons commencé cette étude en partant de l observation qu un mutant sir2 présentait un volume cellulaire augmenté. De manière cohérente, un traitement au nicotinamide (Nam), un inhibiteur de Sir2, reproduit le défaut de taille de sir2 par un mécanisme dépendant de Sir2p. Nous avons alors pu, par une approche d épistasie chimique, identifier 22 mutants non affectés par le traitement de la Nam, parmi ~200 mutants de petite taille. De manière surprenante, 16 de ces 22 mutants de taille insensibles à la NAM, sont affectés dans la biogenèse de la grande sous unité du ribosome (60S). Une drogue capable de bloquer spécifiquement la biogenèse tardive de la 60S, la diazaborine, mime le phénotype de taille des mutants de la 60S, produisant des cellules sauvages plus petites. Un ensemble de ~200 mutants de grande taille a été traité à la diazaborine et leur volume mesuré. Cette approche chimiogénétique nous a permis d identifier 31 mutants insensibles à la diazaborine, incluant swi4 et swi6, deux régulateurs majeurs du cycle cellulaire, critiques pour le contrôle de la taille des cellules. Ces résultats furent confirmés par la construction de double mutants. Ce travail montre qu il est possible d organiser des mutants de taille au sein de voies spécifiques et de définir des relations d épistasie claires entre eux. Nos données indiquent que le contrôle de la taille par cette voie Sir2-60S est indépendant des effets de la ploïdie et du contrôle nutritionnel sur la taille des cellules.Cell size homeostasis implies that specific mechanisms are devoted to coordinating growth and proliferation. It is well established that cell size is affected by nutrient availability and ploidy but the underlying mechanisms are not elucidated. A genome wide search for yeast mutants affected for cell volume homeostasis, conducted in the Tyers lab, revealed that the inactivation of about 400 genes leads to a median cell volume diverging from the isogenic wild-type. The cell size control process is thus a very interesting situation where multiple loci contributing to a complex quantitative trait have been identified but their organisation into distinct pathways and their respective influence remain largely to be elucidated. To address this issue, we started from the observation that a sir2 mutant shows an increased cell size. Consistently, nicotinamide (NAM), a Sir2 inhibitor, mimics the sir2 size defect in a Sir2p-dependent manner. This allowed us to identify among ~200 small size mutants, 22 mutants that were clearly not affected by NAM treatment. Strikingly, 16 out of the 22 NAM unresponsive mutants affected biogenesis of the large ribosomal subunit (named 60S below). Consistently, diazaborine, a drug that blocks the large ribosomal subunit assembly and therefore mimics 60S mutants, rendered wild-type yeast cells smaller. A set of ~200 large mutants were treated with diazaborine and their cell volume was measured. This chemogenetics approach allowed us to identify 31 diazaborine-unresponsive mutants, including swi4 and swi6, two major cell cycle regulators that are critical for cell size control. These results were confirmed by constructing double mutants. This work shows that it is possible to organize cell size mutants in specific pathways and to define clear epistasis relationships between them. Our data indicate that the control of cell size by the Sir2-60S pathway is independent of both the ploidy and the nutritional control of cell size.BORDEAUX2-Bib. électronique (335229905) / SudocSudocFranceF