RESUMEN
Recent studies have suggested a correlation between genotype groups of Brettanomyces bruxellensis and their source of isolation. To further explore this relationship, the objective of this study was to assess metabolic differences in carbon and nitrogen assimilation between different B. bruxellensis strains from three beverages, including beer, wine, and soft drink, using Biolog Phenotype Microarrays. While some similarities of physiology were noted, many traits were variable among strains. Interestingly, some phenotypes were found that could be linked to strain origin, especially for the assimilation of particular α- and ß-glycosides as well as α- and ß-substituted monosaccharides. Based upon gene presence or absence, an α-glucosidase and ß-glucosidase were found explaining the observed phenotypes. Further, using a PCR screen on a large number of isolates, we have been able to specifically link a genomic deletion to the beer strains, suggesting that this region may have a fitness cost for B. bruxellensis in certain fermentation systems such as brewing. More specifically, none of the beer strains were found to contain a ß-glucosidase, which may have direct impacts on the ability for these strains to compete with other microbes or on flavor production.
Asunto(s)
Brettanomyces/genética , Brettanomyces/fisiología , Carbono/metabolismo , Variación Genética , Redes y Vías Metabólicas/genética , Nitrógeno/metabolismo , Cerveza/microbiología , Brettanomyces/clasificación , Brettanomyces/aislamiento & purificación , Bebidas Gaseosas/microbiología , ADN de Hongos/genética , Genómica , Genotipo , Fenotipo , Reacción en Cadena de la Polimerasa , Eliminación de Secuencia , Vino/microbiología , alfa-Glucosidasas/genética , alfa-Glucosidasas/metabolismo , beta-Glucosidasa/genética , beta-Glucosidasa/metabolismoRESUMEN
Mutations in the BRCA1-interacting DEAH helicase Brip1 confer an increased risk of breast cancer. In the present study we aimed to unravel the transcriptional control of Brip1 and to determine its expression levels in a set of 101 primary invasive breast carcinomas. Transcription of Brip1 was found to be cell growth-related and controlled by the E2F/retinoblastoma (Rb) pathway through a conserved E2F-responsive site. Repression of Brip1 expression by the cell growth-inhibiting compound 1alpha,25-dihydroxyvitamin D3 depended on this same E2F-responsive site. In spite of its role as a tumor suppressor, both quantitative reverse transcriptase-PCR analyses and immunohistochemical stainings showed significantly elevated Brip1 expression levels in grade 3 tumors as compared to grade 1 or 2 carcinomas. Furthermore, increased Brip1 transcript levels were found in tumors with an estrogen receptor-negative, progesterone receptor-negative or HER-2-positive status. In conclusion, these data show that Brip1 is a genuine target gene for the E2F/Rb pathway and that elevated expression levels of Brip1 are detected in primary invasive breast carcinomas with unfavorable characteristics.
Asunto(s)
Neoplasias de la Mama/genética , Carcinoma/genética , Proteínas de Unión al ADN/genética , Factores de Transcripción E2F/fisiología , Regulación Neoplásica de la Expresión Génica , ARN Helicasas/genética , Animales , Secuencia de Bases , Factores de Transcripción con Cremalleras de Leucina de Carácter Básico/genética , Sitios de Unión , Neoplasias de la Mama/patología , Carcinoma/patología , Secuencia Conservada , Factores de Transcripción E2F/metabolismo , Proteínas del Grupo de Complementación de la Anemia de Fanconi/genética , Femenino , Humanos , Ratones , Datos de Secuencia Molecular , Invasividad Neoplásica , Homología de Secuencia de Ácido Nucleico , Transcripción Genética , Células Tumorales CultivadasAsunto(s)
Proteínas Bacterianas/genética , Células Epiteliales/microbiología , Análisis por Micromatrices/métodos , Salmonella typhimurium/genética , Salmonella typhimurium/fisiología , Transactivadores/genética , Adhesión Bacteriana , Regulación Bacteriana de la Expresión Génica , Genes Reguladores , Genoma Bacteriano , Análisis por Matrices de Proteínas , Salmonella typhimurium/patogenicidad , Transcripción GenéticaRESUMEN
Duplicated genes can undergo different fates, from nonfunctionalization to subfunctionalization and neofunctionalization. In particular, changes in regulatory sequences affecting the expression domain of genes seem to be responsible for the latter two fates. In this study we used in silico motif detection to show how alterations in the composition of regulatory motifs between paralogous genes in zebrafish and Tetraodon might reflect the functional divergence of duplicates.
Asunto(s)
Peces/genética , Genes Duplicados , Variación Genética , Secuencias Reguladoras de Ácidos Nucleicos/genética , Animales , Secuencia de Bases , Secuencia de Consenso , ADN Intergénico/genética , Bases de Datos Genéticas , Datos de Secuencia Molecular , Homología de Secuencia de Ácido NucleicoRESUMEN
We investigated whether prognostic information is reflected in the expression patterns of ovarian carcinoma samples. RNA obtained from seven FIGO stage I without recurrence, seven platin-sensitive advanced-stage (III or IV), and six platin-resistant advanced-stage ovarian tumors was hybridized on a complementary DNA microarray with 21,372 spotted clones. The results revealed that a considerable number of genes exhibit nonaccidental differential expression between the different tumor classes. Principal component analysis reflected the differences between the three tumor classes and their order of transition. Using a leave-one-out approach together with least squares support vector machines, we obtained an estimated classification test accuracy of 100% for the distinction between stage I and advanced-stage disease and 76.92% for the distinction between platin-resistant versus platin-sensitive disease in FIGO stage III/IV. These results indicate that gene expression patterns could be useful in clinical management of ovarian cancer.
Asunto(s)
Adenocarcinoma/genética , Adenocarcinoma/patología , Neoplasias Ováricas/genética , Neoplasias Ováricas/patología , Adenocarcinoma/tratamiento farmacológico , Adulto , Anciano , Anciano de 80 o más Años , Antineoplásicos/uso terapéutico , Resistencia a Antineoplásicos , Femenino , Perfilación de la Expresión Génica , Humanos , Persona de Mediana Edad , Estadificación de Neoplasias , Análisis de Secuencia por Matrices de Oligonucleótidos , Neoplasias Ováricas/tratamiento farmacológico , Compuestos de Platino/uso terapéutico , Valor Predictivo de las PruebasRESUMEN
Quorum sensing (QS) in Gram-negative bacteria is generally assumed to be mediated by N-acyl-homoserine lactone molecules while Gram-positive bacteria make use of signaling peptides. We analyzed the occurrence in Gram-negative bacteria of peptides and transporters that are involved in quorum sensing in Gram-positive bacteria. Many class II bacteriocins and inducing factors produced by lactic acid bacteria (LAB) and competence stimulating peptides (CSPs) synthesized by streptococci are processed by their cognate ABC-transporters during their secretion. During transport, a conserved leader sequence, termed the double-glycine motif (GG-motif), is cleaved off by the N-terminal domain of the transporter, which belongs to the Peptidase C39 protein family. Several peptides containing a GG-motif were recently described in Gram-negative bacteria (Trends Microbiol 2001;9:164-8). To screen for additional putative GG-motif containing peptides, an in silico strategy based on MEME, HMMER2.2 and Wise2 was designed. Using a curated training set, a motif model of the leader peptide was built and used to screen over 120 fully sequenced bacterial genomes. The screening methodology was applied at the nucleotide level as probably many small peptide genes have not been annotated and may be absent from the non-redundant databases. It was found that 33% of the screened genomes of Gram-negative bacteria contained one or more transporters carrying a Peptidase C39 domain, compared to 44% of the genomes of Gram-positive bacteria. The transporters can be subdivided into four classes on the basis of their domain organization. Genes coding for putative peptides containing 23-142 amino acids and a GG-motif were found in close association with genes coding for Peptidase C39 domain containing proteins. These peptides show structural similarity to bacteriocins and peptide pheromones of Gram-positive bacteria. The possibility of signal transduction based on peptide signaling in Gram-negative bacteria is discussed.
Asunto(s)
Bacteriocinas/química , Genoma Bacteriano , Glicina/química , Bacterias Gramnegativas/fisiología , Péptidos/química , Secuencias de Aminoácidos , Secuencia de Aminoácidos , Proteínas Bacterianas/biosíntesis , Bacteriocinas/metabolismo , Transporte Biológico , Comunicación Celular , Biología Computacional , Modelos Biológicos , Datos de Secuencia Molecular , Filogenia , Plásmidos/metabolismo , Estructura Terciaria de Proteína , Homología de Secuencia de Aminoácido , Transducción de Señal , Programas InformáticosRESUMEN
MOTIVATION: Transcriptome analysis allows detection and clustering of genes that are coexpressed under various biological circumstances. Under the assumption that coregulated genes share cis-acting regulatory elements, it is important to investigate the upstream sequences controlling the transcription of these genes. To improve the robustness of the Gibbs sampling algorithm to noisy data sets we propose an extension of this algorithm for motif finding with a higher-order background model. RESULTS: Simulated data and real biological data sets with well-described regulatory elements are used to test the influence of the different background models on the performance of the motif detection algorithm. We show that the use of a higher-order model considerably enhances the performance of our motif finding algorithm in the presence of noisy data. For Arabidopsis thaliana, a reliable background model based on a set of carefully selected intergenic sequences was constructed. AVAILABILITY: Our implementation of the Gibbs sampler called the Motif Sampler can be used through a web interface: http://www.esat.kuleuven.ac.be/~thijs/Work/MotifSampler.html. CONTACT: gert.thijs@esat.kuleuven.ac.be; yves.moreau@esat.kuleuven.ac.be
Asunto(s)
Algoritmos , Simulación por Computador , Modelos Genéticos , Probabilidad , Regiones Promotoras Genéticas , Arabidopsis/genética , ADN Intergénico , Expresión Génica , Transcripción GenéticaRESUMEN
An Azospirillum brasilense Sp7 strain containing a plasmid-borne translational cytN-gusA fusion was grown in a continuous culture to quantitatively evaluate the influence of extracellular signals (such as O(2)) on expression of the cytNOQP operon. The dissolved oxygen concentration was shifted at regular time intervals before the steady state was reached. The measured beta-glucuronidase activity was used to monitor cytN gene expression. However, as the beta-glucuronidase activity in the experimental setup not only depended on altered transcription of the hybrid gene when the signal was varied but was also influenced by cellular accumulation, degradation, and dilution of the hybrid fusion protein, a mathematical method was developed to describe the intrinsic properties of the dynamic bioprocess. After identification and validation of the mathematical model, the apparent specific rate of expression of the fusion, which was independent of the experimental setup, could be deduced from the model and used to quantify gene expression regulated by extracellular environmental signals. In principle, this approach can be generalized to assess the effects of external signals on bacterial gene expression.
Asunto(s)
Azospirillum brasilense/genética , Regulación Bacteriana de la Expresión Génica , Genes Reporteros , Glucuronidasa/genética , Modelos Biológicos , Azospirillum brasilense/crecimiento & desarrollo , Azospirillum brasilense/metabolismo , Medios de Cultivo , Complejo IV de Transporte de Electrones/genética , Complejo IV de Transporte de Electrones/metabolismo , Glucuronidasa/metabolismo , Oxígeno/farmacología , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/metabolismoRESUMEN
Disruption of an open reading frame (ORF) of 840 bp (280 amino acids; ORF280) in an Azospirillum brasilense Tn5 mutant resulted in a pleiotrophic phenotype. Besides an enhanced N(2)-fixing capacity and altered expression pattern of a nifH-gusA fusion, growth on the charged polar amino acids glutamate and arginine was severely affected. ORF280, similar to previously identified ORFs present in Bradyrhizobium japonicum (ORF277), Paracoccus denitrificans (ORF278) and Rhodobacter capsulatus (ORF277), exhibits in its C-terminus a significant similarity with the recently defined family of universal stress proteins.
Asunto(s)
Azospirillum brasilense/genética , Elementos Transponibles de ADN , Fijación del Nitrógeno/genética , Nitrogenasa/genética , Sistemas de Lectura Abierta/genética , Oxidorreductasas , Secuencia de Aminoácidos , Azospirillum brasilense/metabolismo , Clonación Molecular , ADN Bacteriano/análisis , Glucuronidasa/metabolismo , Datos de Secuencia Molecular , Mutación , Fijación del Nitrógeno/fisiología , Nitrogenasa/metabolismo , Mapeo Físico de Cromosoma , Proteínas Recombinantes de Fusión/metabolismo , Análisis de Secuencia de ADNRESUMEN
Spectral analysis indicated the presence of a cytochrome cbb3 oxidase under microaerobic conditions in Azospirillum brasilense Sp7 cells. The corresponding genes (cytNOQP) were isolated by using PCR. These genes are organized in an operon, preceded by a putative anaerobox. The phenotype of an A. brasilense cytN mutant was analyzed. Under aerobic conditions, the specific growth rate during exponential phase (mu(e)) of the A. brasilense cytN mutant was comparable to the wild-type specific growth rate (m(e) of approximately 0.2 h-1). In microaerobic NH4+-supplemented conditions, the low respiration of the A. brasilense cytN mutant affected its specific growth rate (mu(e) of approximately 0.02 h-1) compared to the wild-type specific growth rate (mu(e) of approximately 0.2 h-1). Under nitrogen-fixing conditions, both the growth rates and respiration of the wild type were significantly diminished in comparison to those under NH4+-supplemented conditions. Differences in growth rates and respiration between the wild type and the A. brasilense cytN mutant were less pronounced under these nitrogen-fixing conditions (mu(e) of approximately 0.03 h-1 for the wild type and 0.02 h-1 for the A. brasilense cytN mutant). The nitrogen-fixing capacity of the A. brasilense cytN mutant was still approximately 80% of that determined for the wild-type strain. This leads to the conclusion that the A. brasilense cytochrome cbb3 oxidase is required under microaerobic conditions, when a high respiration rate is needed, but that under nitrogen-fixing conditions the respiration rate does not seem to be a growth-limiting factor.
Asunto(s)
Azospirillum brasilense/enzimología , Azospirillum brasilense/crecimiento & desarrollo , Complejo IV de Transporte de Electrones/fisiología , Oxidorreductasas/fisiología , Aerobiosis , Azospirillum brasilense/genética , Secuencia de Bases , Membrana Celular/enzimología , ADN Bacteriano , Complejo IV de Transporte de Electrones/genética , Espectrometría de Masas , Datos de Secuencia Molecular , Mutagénesis , Fijación del Nitrógeno , Oxidorreductasas/genética , Fenotipo , Análisis de Secuencia de ADNRESUMEN
Although HIV is still seen by many as a disease affecting younger adults, it is known that at least 11% of individuals with AIDS in Britain are over the age of 50. It is likely that this older age representation will continue and increase and whilst much is known about the psychological wellbeing of younger individuals with HIV, it is crucial that we consider the differing experiences and needs of older adults with the disease. Fifty-two adults with HIV over the age of 54 were referred to the Department of Psychological Medicine between June 1990 and December 1996. Data for these patients were compared with corresponding information for a random sample of younger patients with HIV. The older adults were found to differ significantly from the younger individuals on a variety of social, psychological and medical variables including social isolation, employment worries, sexuality, previous psychiatric history and stage of HIV at referral. It seems that current HIV services may actually alienate a significant proportion of potential users through not being sensitive to the needs and views of these older individuals. This must be addressed by policy makers and practitioners to ensure that psychiatric and psychological services become more acceptable and accessible to the older adult.