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1.
Phys Rev Lett ; 131(11): 116902, 2023 Sep 15.
Artículo en Inglés | MEDLINE | ID: mdl-37774304

RESUMEN

Optically active spin defects in hexagonal boron nitride (hBN) are promising quantum systems for the design of two-dimensional quantum sensing units offering optimal proximity to the sample being probed. In this Letter, we first demonstrate that the electron spin resonance frequencies of boron vacancy centers (V_{B}^{-}) can be detected optically in the limit of few-atomic-layer thick hBN flakes despite the nanoscale proximity of the crystal surface that often leads to a degradation of the stability of solid-state spin defects. We then analyze the variations of the electronic spin properties of V_{B}^{-} centers with the hBN thickness with a focus on (i) the zero-field splitting parameters, (ii) the optically induced spin polarization rate and (iii) the longitudinal spin relaxation time. This Letter provides important insights into the properties of V_{B}^{-} centers embedded in ultrathin hBN flakes, which are valuable for future developments of foil-based quantum sensing technologies.

2.
Nat Commun ; 13(1): 4347, 2022 Jul 27.
Artículo en Inglés | MEDLINE | ID: mdl-35896526

RESUMEN

Spin defects in hexagonal boron nitride (hBN) are promising quantum systems for the design of flexible two-dimensional quantum sensing platforms. Here we rely on hBN crystals isotopically enriched with either 10B or 11B to investigate the isotope-dependent properties of a spin defect featuring a broadband photoluminescence signal in the near infrared. By analyzing the hyperfine structure of the spin defect while changing the boron isotope, we first confirm that it corresponds to the negatively charged boron-vacancy center ([Formula: see text]). We then show that its spin coherence properties are slightly improved in 10B-enriched samples. This is supported by numerical simulations employing cluster correlation expansion methods, which reveal the importance of the hyperfine Fermi contact term for calculating the coherence time of point defects in hBN. Using cross-relaxation spectroscopy, we finally identify dark electron spin impurities as an additional source of decoherence. This work provides new insights into the properties of [Formula: see text] spin defects, which are valuable for the future development of hBN-based quantum sensing foils.

4.
Virus Res ; 159(2): 115-23, 2011 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-21605606

RESUMEN

Watermelon mosaic virus (WMV, genus Potyvirus, family Potyviridae) was reported for the first time in France in 1974, and it is now the most prevalent virus in cucurbit crops. In 2000, new strains referred as 'emerging' (EM) strains were detected in South-eastern France. EM strains are generally more severe and phylogenetically distinct from those previously reported in this country and referred as 'classic' (CL) strains. Since 2000, EM strains have been progressively replacing CL strains in several areas where they co-exist. In order to explain this rapid shift in virus populations, the biological properties of a set of 17 CL and EM WMV isolates were compared. No major differences were observed when comparing a limited host range including 48 different plant species or cultivars. Only two species were differential; Chenopodium quinoa was systemically infected by CL and not by EM isolates whereas Ranunculus sardous was systemically infected by EM and not by CL isolates. A considerable variability was observed in aphid transmission efficiencies but this could not be correlated to the CL or EM types. Two subsets of five isolates of each group were used to compare aphid transmission efficiencies from single and double (CL-EM) infections using six different cucurbit and non-cucurbit hosts. EM isolates were generally better transmitted from mixed CL-EM infections than CL isolates and CL transmission rates were significantly lower from double than from single infections. Cross-protection was only partial between CL and EM strains leading to frequent double infections, and only a slight asymmetry was observed in cross-protection efficiencies. Since double infections occur very commonly in fields, the preferential transmission of EM from mixed CL-EM infections could be one of the factors leading to the displacement of CL isolates by EM isolates.


Asunto(s)
Cucurbita/virología , Variación Genética , Filogeografía , Enfermedades de las Plantas/virología , Potyvirus/genética , Potyvirus/patogenicidad , Animales , Áfidos/virología , Protección Cruzada , Vectores de Enfermedades , Francia , Especificidad del Huésped , Potyvirus/clasificación
5.
Virus Res ; 141(2): 190-200, 2009 May.
Artículo en Inglés | MEDLINE | ID: mdl-19152815

RESUMEN

Cucurbit viruses are involved in complex and changing pathosystems in France, with new virus strains or species regularly reported. Zucchini yellow mosaic virus (ZYMV) is an archetypal emerging virus that was reported in France in 1979. It has since caused sporadic but occasionally very severe economic losses and its epidemiology still remains poorly understood. Partial sequencing of the viral genome has been used to characterize ZYMV isolates that occurred over a 29-year period in experimental plots at Montfavet, France (n=227), or that were received through a national survey for cucurbit viruses conducted in France from 2004 to 2007 (n=198). A total of 34 haplotypes were differentiated belonging to five molecular groups, three including isolates already described in France and two corresponding to isolates that emerged in France within the last 5 years. Comparison of haplotypes found at one location during successive years revealed contrasting situations. When they were either the same or closely related haplotypes, this suggested the availability of overwintering hosts, whereas when they belonged to different molecular groups this indicated shifts in viral populations with possible new introductions. The contribution of molecular epidemiology in tracing the origin of ZYMV in the French West Indies is also reviewed.


Asunto(s)
Cucurbita/virología , Enfermedades de las Plantas/virología , Potyvirus/genética , Francia , Epidemiología Molecular , Datos de Secuencia Molecular , Filogenia , Potyvirus/clasificación , Potyvirus/aislamiento & purificación
6.
Ann Dermatol Venereol ; 136(1): 32-6, 2009 Jan.
Artículo en Francés | MEDLINE | ID: mdl-19171227

RESUMEN

INTRODUCTION: Palmar or plantar circumscribed hypokeratosis is a remarkable clinicopathologic entity described in 2002. It consists in a well demarcated decrease in thickness of the stratum corneum, that can be clinically mistaken for Bowen's disease or porokeratosis. We present a classical plantar localisation and a more original case on the dorsum of the finger, together with a microscopic and immunohistochemical study. CASE REPORTS: Case 1. A 65-year-old man was seen 15 years after a first consultation for a well demarcated 1.5cm erythematous lesion localised on the border of his left foot. The biopsy, then misinterepreted as keratosis sulcata, was reviewed. It showed a sudden and well demarcated decrease in thickness of the stratum corneum, overlying a slightly acanthotic epidermis, associated with dilated capillaries in the papillary dermis. HPV immunostaining was negative. Case 2. A 75-year-old woman had a well demarcaated erythematous lesion of the dorsum of her right index finger, lasting for months without significant evolution. A first biopsy showed pale and haloed keratinocytes that could be interpreted as koilocytes. She was therefore treated by cryotherapy, 5-fluro-uracile and imiquimod, that proved unsuccessful. A second biopsy showed a sudden and major decrease in thickness of the stratum corneum, overlying an area containing a few pale keratinocytes with perinuclear halo. HPV immunostaing was negative and Ki67 positive cells were slightly decreased in number when compared to lateral normal skin. DISCUSSION: Our first case is typical of plantar hypokeratosis characterised by its long evolution, typical semiology and well demarcated anomaly of the stratum corneum. Our second case is original as it shows that the disease can also affect the dorsum of the fingers. Acral circumscribed hypokeratosis is therefore a better name for this condition. We did not find any arguments in favor of a viral cause or an increased proliferation of keratinocytes. As often described in other cases, the lesion can remain unchanged for decades, which confirm its benign evolution. Topical treatments are generally ineffective. The pathogenesis of this localised hypokeratosis remains mysterious.


Asunto(s)
Dermatosis del Pie/patología , Dermatosis de la Mano/patología , Queratosis/patología , Anciano , Humanos , Masculino , Persona de Mediana Edad
7.
Phytopathology ; 97(11): 1484-90, 2007 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-18943519

RESUMEN

ABSTRACT Decision rule models are considered to be one of the main cornerstones of the implementation of integrated pest management (IPM) programs. Even if the need for such programs to offer cost advantages over conventional strategies is a major incentive for IPM adoption, few studies focus on this financial dimension. In this article, a modeling approach of the response of a pathosystem to a disease control method and of the predictive performance of decision rules is used to explore how some basic factors act on the likelihood of adoption of decision rule models strategies (such as using an IPM system) over systematic strategies (such as systematic-spraying and never-spraying strategies). Even if the average cost of using the decision rule strategies is always lower than the average cost of systematic strategies in several different scenarios, the models developed here showed strong effects of different pathosystems and decision rules on financial benefits. The number of production situations where decision rules are of interest is highly correlated with their accuracy. However, because of the inescapable trade-offs between decision rule accuracy and limiting factors such as its user-friendly characteristics, the use of decision rules is unlikely to reduce costs to <70% of the costs of systemic strategies. In more general terms, this study provides quantitative guidelines on the financial advantage that decision rules can offer in plant protection as well as a better understanding of their potential usefulness.

8.
J Econ Entomol ; 97(3): 1137-41, 2004 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-15279302

RESUMEN

The melon fly, Bactrocera cucurbitae (Coquillett) (Diptera: Tephritidae), is the most damaging pest of cucurbits in Reunion Island. The influence of adding borax and modifying pH on the effectiveness of different food attractants for both sexes of the melon fly is analyzed by a release-recapture method in field cages. Adding borax to protein hydrolysates Nulure and Buminal strongly reduced their attractiveness for B. cucurbitae. Acidification of 5% Buminal solution (from pH 6 to pH 3) doubled its attractiveness for melon fly. Conversely, Torula yeast at pH 10.5 was significantly more attractive than the standard Torula yeast at pH 9 (28% of captured flies compared with 17%). However, a further pH increase of the yeast solution does not improve its attractiveness. The results are discussed in relation to other studies on pH modification of various baits for Tephritidae.


Asunto(s)
Boratos/administración & dosificación , Control de Insectos/métodos , Feromonas , Tephritidae , Animales , Cucurbita , Concentración de Iones de Hidrógeno
9.
J Econ Entomol ; 96(1): 231-8, 2003 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-12650367

RESUMEN

The relative attractiveness of six commercially available protein hydrolysates and the influence of their concentration were evaluated in field cages by a release-capture method of lab-reared melon fly adults, Bactrocera cucurbitae (Coquillett). Buminal, Corn Steepwater, Hym-Lure, Pinnacle, Nulure, and SolBait were tested for both sexes of the melon fly. The tested products exhibited clear differences in attractiveness. SolBait was the most effective protein hydrolysate. Pinnacle and Corn Steepwater also gave promising results. A general tendency for an increase in effectiveness with increasing concentration within the range 0.5 to 10% was shown. This study will allow pest control practitioners to choose more effective attractants for use in bait sprays to control the melon fly thus reducing the intensive use of insecticides currently practiced in Reunion island and enabling the development of Integrated pest management (IPM) methods for cucurbit crops.


Asunto(s)
Dípteros/fisiología , Control de Insectos/métodos , Hidrolisados de Proteína , Animales , Femenino , Masculino , Reunión
10.
Phytopathology ; 93(10): 1217-22, 2003 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-18944319

RESUMEN

ABSTRACT Barley yellow dwarf virus (BYDV) damage to winter cereals and population dynamics of the aphid Rhopalosiphum padi during fall were monitored in fields during 10 years at various locations in the northern half of France. Logistic regression was used to examine whether a simple risk probability algorithm based only on the autumnal population dynamics of R. padi can accurately predict yield losses caused by BYDV and, therefore, the need for insecticide treatment. Results showed that the area under the curve of the percentage of plants infested by R. padi during autumn was highly significantly related to BYDV yield losses. Then, a cost/benefit analysis was performed to estimate the optimal decision threshold resulting in the lowest annual average costs of BYDV damage and control. A "model use" strategy allowed a reduction in the annual average costs of BYDV disease and control of up to 36% when compared with a "prophylactic spraying" strategy. The optimal decision threshold was highly sensitive to variation in disease prevalence. This property was used to propose an easy way to adapt the model to any production situation through the determination of the most accurate decision threshold.

11.
Mol Genet Genomics ; 266(6): 979-87, 2002 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-11862492

RESUMEN

Sunflower (Helianthus annuus L.) is an economically important oil seed crop with an estimated genome size of 3000 Mb. We have constructed a bacterial artificial chromosome (BAC) library for sunflower, which represents an estimated 4- to 5-fold coverage of the genome. Nuclei isolated from young leaves were used as a source of high-molecular-weight DNA and a partial restriction endonuclease digestion protocol was used to cleave the DNA. A random sample of 60 clones indicated an average insert size of 80 kb, implying a 95% probability of recovering any specific sequence of interest. The library was screened with chloroplast DNA probes. Only 0.1% of the clones were identified to be of chloroplast origin, indicating that contamination with organellar DNAs is very low. The utility of the library was evaluated by screening for the presence of genes for putative transmembrane receptors sharing epidermal growth factor (EGF) and integrin-like domains. First, a homologous sunflower EST (HaELP1) was obtained by degenerate RT-PCR cloning, using Arabidopsis thaliana genes (AtELP) as a source of consensus sequences. Three different BACs yielded positive hybridization signals when HaELP1 was used as a probe. BAC subcloning and sequencing demonstrated the presence of two different loci putatively homologous to genes for transmembrane proteins with EGF- and integrin-like domains from sunflower. This work demonstrates the suitability of the library for homology map-based cloning of sunflower genes and physical mapping of the sunflower genome.


Asunto(s)
Biblioteca de Genes , Genes de Plantas/genética , Helianthus/genética , Proteínas de la Membrana/genética , Secuencia de Aminoácidos , Secuencia de Bases , Cromosomas Artificiales Bacterianos/genética , Clonación Molecular , ADN Complementario/química , ADN Complementario/genética , Datos de Secuencia Molecular , Receptores de Superficie Celular/genética , Alineación de Secuencia , Análisis de Secuencia de ADN , Homología de Secuencia de Aminoácido
13.
J Exp Bot ; 52(362): 1857-64, 2001 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-11520874

RESUMEN

The identification of QTL for several physiological traits in sunflower is described. Traits related to photosynthesis (leaf chlorophyll concentration, net photosynthesis and internal CO(2) concentration) and water status (stomatal conductance, transpiration, predawn leaf water potential, and relative water content) were evaluated in a population of recombinant inbred lines under greenhouse conditions. Narrow-sense heritabilities were low to average. Using an AFLP linkage map, 19 QTL were detected explaining 8.8-62.9% of the phenotypic variance for each trait. Among these, two major QTL for net photosynthesis were identified on linkage group IX. One QTL co-location was found on linkage group VIII for stomatal movements and water status. Coincident locations for QTL regulating photosynthesis, transpiration and leaf water potential were described on linkage group XIV. These results lead to the first description of the organization of genomic regions related to yield in sunflower.


Asunto(s)
Helianthus/genética , Fotosíntesis/genética , Carácter Cuantitativo Heredable , Dióxido de Carbono/metabolismo , Clorofila/metabolismo , Mapeo Cromosómico , Ambiente Controlado , Genes de Plantas , Marcadores Genéticos , Helianthus/fisiología , Presión Osmótica , Fotosíntesis/fisiología , Epidermis de la Planta/fisiología , Hojas de la Planta/genética , Hojas de la Planta/fisiología , Transpiración de Plantas , Recombinación Genética , Agua/metabolismo
14.
EMBO J ; 19(13): 3408-17, 2000 Jul 03.
Artículo en Inglés | MEDLINE | ID: mdl-10880453

RESUMEN

Recombination events between non-identical sequences most often involve heteroduplex DNA intermediates that are subjected to mismatch repair. The well-characterized long-patch mismatch repair process, controlled in eukaryotes by bacterial MutS and MutL orthologs, is the major system involved in repair of mispaired bases. Here we present evidence for an alternative short-patch mismatch repair pathway that operates on a broad spectrum of mismatches. In msh2 mutants lacking the long-patch repair system, sequence analysis of recombination tracts resulting from exchanges between similar but non-identical (homeologous) parental DNAs showed the occurrence of short-patch repair events that can involve <12 nucleotides. Such events were detected both in mitotic and in meiotic recombinants. Confirming the existence of a distinct short-patch repair activity, we found in a recombination assay involving homologous alleles that closely spaced mismatches are repaired independently with high efficiency in cells lacking MSH2 or PMS1. We show that this activity does not depend on genes required for nucleotide excision repair and thus differs from the short-patch mismatch repair described in Schizosaccharomyces pombe.


Asunto(s)
Disparidad de Par Base , Reparación del ADN , Saccharomyces cerevisiae/genética , Secuencia de Bases , ADN de Hongos , Datos de Secuencia Molecular , Mutación
15.
Nat Genet ; 25(2): 192-4, 2000 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-10835635

RESUMEN

DNA helicases are involved in many aspects of DNA metabolism, including transcription, replication, recombination and repair. In the yeast Saccharomyces cerevisiae, the absence of the Sgs1 helicase results in genomic instability and accelerated ageing. In human cells, mutations in orthologues of SGS1 lead to Bloom (BS), Werner (WS) or Rothmund-Thomson (RTS) syndromes, which are rare, autosomal recessive diseases characterized by genetic instability associated with cancer predisposition. Although data concerning these human diseases are accumulating, there is still no clear idea of the function of the proteins involved. Here we show that sgs1Delta mutants are deficient in DNA repair and are defective for induced recombination events that involve homologous chromosomes. The role of homologous recombination is further evidenced in haploid cells in which both Sgs1p and Srs2p are absent. Yeast SRS2 encodes another DNA helicase involved in the maintenance of genome integrity. Our data suggest that some defects observed in BS, WS or RTS are the consequence of unrestrained recombination.


Asunto(s)
ADN Helicasas/metabolismo , Proteínas Fúngicas/metabolismo , Recombinación Genética/genética , Proteínas de Saccharomyces cerevisiae , Saccharomyces cerevisiae/enzimología , Saccharomyces cerevisiae/genética , Homología de Secuencia de Ácido Nucleico , Síndrome de Bloom/genética , División Celular/genética , División Celular/efectos de la radiación , Daño del ADN/genética , Daño del ADN/efectos de la radiación , ADN Helicasas/deficiencia , ADN Helicasas/genética , Reparación del ADN/genética , Replicación del ADN , Proteínas de Unión al ADN/genética , Proteínas de Unión al ADN/fisiología , Proteínas Fúngicas/genética , Rayos gamma , Genes Fúngicos/genética , Genes Fúngicos/fisiología , Genes Letales/genética , Haploidia , Humanos , Mutación/genética , Recombinasa Rad51 , Tolerancia a Radiación/genética , RecQ Helicasas , Recombinación Genética/efectos de la radiación , Síndrome Rothmund-Thomson/genética , Saccharomyces cerevisiae/citología , Saccharomyces cerevisiae/efectos de la radiación , Intercambio de Cromátides Hermanas/genética , Intercambio de Cromátides Hermanas/efectos de la radiación , Esporas Fúngicas/citología , Esporas Fúngicas/enzimología , Esporas Fúngicas/genética , Esporas Fúngicas/efectos de la radiación , Rayos Ultravioleta , Síndrome de Werner/genética
16.
J Chromatogr B Biomed Sci Appl ; 734(2): 267-76, 1999 Nov 12.
Artículo en Inglés | MEDLINE | ID: mdl-10595724

RESUMEN

This paper describes a GC-MS method (SIM mode) for the analysis of perfluorooctyl bromide (perflubron, I) in rat blood. The chromatographic separation was performed by injection in the split mode using a CP-select 624 CB capillary column. Following destruction of the emulsion by addition of ethanol, the analytical procedure involves a liquid-liquid extraction with 1,1,2-trichlorotrifluoroethane. The bis(F-butyl)ethene (II) was used as internal standard. Observed retention times were 3.22 min for I and 2.32 min for II. Two calibration curves were used; linear detection responses were obtained for concentrations ranging from 0.009 to 0.9 mg/ml and from 0.9 to 13.5 mg/ml. The extraction efficiency averaged 50% for I and 93% for II. Precision ranged from 0.7 to 14%, and accuracy was between 91 and 109%. The limit of quantification was 9 microg/ml. The method validation results indicate that the performance characteristics of the method fulfilled the requirements for assay method for use in pharmacokinetic studies.


Asunto(s)
Fluorocarburos/sangre , Cromatografía de Gases y Espectrometría de Masas/métodos , Animales , Fluorocarburos/farmacocinética , Semivida , Hidrocarburos Bromados , Cinética , Masculino , Control de Calidad , Ratas , Ratas Wistar , Sensibilidad y Especificidad
17.
Yeast ; 15(14): 1529-39, 1999 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-10514570

RESUMEN

We have carried out the systematic disruption of six ORFs on chromosome XV, of Saccharomyces cerevisiae using the long flanking homology technique to replace each with the KanMX cassette; we have also constructed plasmids containing replacement cassettes and cognate clones for each ORF. Disruption of three of the ORFs-YOL117w, YOL114c, and YOL112w (also known as MSB4)-does not result in any noteworthy phenotype with respect to temperature or nutritional requirements, but yol112w mutants with an additional disruption of YNL293w, which encodes a protein similar to Yol112w, exhibit a slow growth phenotype. The protein specified by YOL114c shares similarity with the human DS-1 protein. Disruption of YOL115w confers slow growth, cold sensitivity and poor sporulation; this ORF has been described elsewhere as TRF4, which encodes a topoisomerase I-related protein. Cells with disruptions of YOL111c, whose product is weakly similar to the human ubiquitin-like protein GdX, are slightly impaired in mating. Mutants disrupted for YOL072w, the predicted product of which is unrelated to any protein of known function, grow slowly, are cold-sensitive and sporulate with reduced efficiency.


Asunto(s)
Proteínas Cromosómicas no Histona/genética , Cromosomas Fúngicos , ADN Polimerasa Dirigida por ADN , Proteínas Nucleares , Sistemas de Lectura Abierta , Proteínas de Saccharomyces cerevisiae , Saccharomyces cerevisiae/genética
18.
Yeast ; 15(12): 1255-67, 1999 Sep 15.
Artículo en Inglés | MEDLINE | ID: mdl-10487928

RESUMEN

In the context of the EUROFAN project, we have carried out the systematic disruption of seven ORFs on chromosome IV of Saccharomyces cerevisiae using the long flanking homology technique to replace each ORF with the KanMX cassette. Targeted disruption of YDL057w, YDL012c, or YDL010w with YDL009c (the two ORFs overlap) confers no overt defects in haploid growth on a variety of media at different temperatures, in mating, or in the sporulation of diploids homozygous for the disruption. By contrast, YDL008w and YDL003w disruptants are non-viable. The product of YDL008w (elsewhere identified as APC11) is a component of the anaphase promoting complex. YDL003w (also termed MCD1) is a homologue of Schizosaccharomyces pombe rad21, an essential gene implicated in DNA double-strand break repair and nuclear organization in fission yeast. In budding yeast, this ORF has been shown by several laboratories to encode a protein involved in sister chromatid cohesion and chromosome condensation. The remaining ORF, YDL005c (also termed MED2), encodes a component of the transcriptional activator complex known as Mediator. Disruption of YDL005c confers a modest slow growth phenotype on rich medium and a more severe phenotype on minimal medium, aberrant cellular morphology, and mating defects; diploids homozygous for the disruption cannot sporulate.


Asunto(s)
Cromosomas Fúngicos/genética , Genes Fúngicos/genética , Mutagénesis Insercional , Sistemas de Lectura Abierta/genética , Saccharomyces cerevisiae/genética , División Celular , Segregación Cromosómica , Medios de Cultivo , Reparación del ADN , Proteínas Fúngicas/química , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Genes Esenciales/genética , Genes Esenciales/fisiología , Genes Fúngicos/fisiología , Prueba de Complementación Genética , Peso Molecular , Mutación/genética , Fenotipo , Saccharomyces cerevisiae/crecimiento & desarrollo , Saccharomyces cerevisiae/fisiología , Homología de Secuencia de Aminoácido , Esporas Fúngicas/genética , Esporas Fúngicas/fisiología , Temperatura , Activación Transcripcional
19.
EMBO J ; 18(6): 1701-11, 1999 Mar 15.
Artículo en Inglés | MEDLINE | ID: mdl-10075939

RESUMEN

Yeast cells mutant for TOP3, the gene encoding the evolutionary conserved type I-5' topoisomerase, display a wide range of phenotypes including altered cell cycle, hyper-recombination, abnormal gene expression, poor mating, chromosome instability and absence of sporulation. In this report, an analysis of the role of TOP3 in the meiotic process indicates that top3Delta mutants enter meiosis and complete the initial steps of recombination. However, reductional division does not occur. Deletion of the SPO11 gene, which prevents recombination between homologous chromosomes in meiosis I division, allows top3Delta mutants to form viable spores, indicating that Top3 is required to complete recombination successfully. A topoisomerase activity is involved in this process, since expression of bacterial TopA in yeast top3Delta mutants permits sporulation. The meiotic block is also partially suppressed by a deletion of SGS1, a gene encoding a helicase that interacts with Top3. We propose an essential role for Top3 in the processing of molecules generated during meiotic recombination.


Asunto(s)
ADN-Topoisomerasas de Tipo I/genética , ADN-Topoisomerasas de Tipo I/metabolismo , Recombinación Genética , Saccharomyces cerevisiae/fisiología , Secuencia de Aminoácidos , Ciclo Celular/genética , Núcleo Celular/ultraestructura , Secuencia Conservada , Daño del ADN , ADN de Hongos/genética , Evolución Molecular , Eliminación de Gen , Genotipo , Meiosis , Mutagénesis , Saccharomyces cerevisiae/enzimología , Saccharomyces cerevisiae/genética , Esporas Fúngicas
20.
Eur J Neurosci ; 10(5): 1631-43, 1998 May.
Artículo en Inglés | MEDLINE | ID: mdl-9751136

RESUMEN

To study a step of the very complex processes of the formation of the neuromuscular junction (NMJ), we have analysed the clustering of acetylcholine receptors (AChR) and acetylcholinesterase (AChE) in myotubes cultured in various conditions. On the surface of rat myotubes cultured in the presence of spinal cord cells from embryonic rat, numerous AChE clusters appeared. Such clusters are always co-localized with AChR clusters, but the reverse is not true: the number of AChR clusters largely exceeds that of AChE clusters. Very few AChE clusters formed when such co-cultures were treated with monoclonal antibodies (mAbs) against the main immunogenic region (MIR) of the AChR, which provoke internalization and degradation of the AChRs of the muscular membrane. The total levels of AChE and proportions of molecular forms were unaffected. We also used non-innervated myotubes in which addition of agrin, a protein normally synthesized by motoneurons, transported to nerve terminals and inserted into the synaptic basal lamina, induces the formation of small clusters of AChE. When added to rat myotubes devoid of membrane AChR, agrin-induced AChE clusters did not form. Finally, we analysed the capacity of the variant of the C2 mouse muscle cell line deficient in AChR (1R-) to form clusters of AChE in co-cultures with spinal cord cells from rat: no formation of AChE clusters could be observed. In all these different systems of cultures, the conditions which prevented clustering of AChR (anti-AChR antibodies, deficiency of the variant C2 cell line) also suppressed AChE clustering. We concluded that clustering of AChR is a prerequisite for clustering of AChE, so that NMJ formation implies the sequential accumulation of these two components.


Asunto(s)
Acetilcolinesterasa/metabolismo , Unión Neuromuscular/fisiología , Receptores Colinérgicos/metabolismo , Sinapsis/fisiología , Animales , Anticuerpos Monoclonales , Especificidad de Anticuerpos , Técnicas de Cocultivo , Femenino , Laminina , Masculino , Ratas , Ratas Sprague-Dawley , Coloración y Etiquetado
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