Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 15 de 15
Filtrar
1.
Cell ; 155(7): 1556-67, 2013 Dec 19.
Artículo en Inglés | MEDLINE | ID: mdl-24360277

RESUMEN

Gene expression has to withstand stochastic, environmental, and genomic perturbations. For example, in the latter case, 0.5%-1% of the human genome is typically variable between any two unrelated individuals. Such diversity might create problematic variability in the activity of gene regulatory networks and, ultimately, in cell behaviors. Using multigenerational selection experiments, we find that for the Drosophila proneural network, the effect of genomic diversity is dampened by miR-9a-mediated regulation of senseless expression. Reducing miR-9a regulation of the Senseless transcription factor frees the genomic landscape to exert greater phenotypic influence. Whole-genome sequencing identified genomic loci that potentially exert such effects. A larger set of sequence variants, including variants within proneural network genes, exhibits these characteristics when miR-9a concentration is reduced. These findings reveal that microRNA-target interactions may be a key mechanism by which the impact of genomic diversity on cell behavior is dampened.


Asunto(s)
Proteínas de Drosophila/metabolismo , Drosophila melanogaster/genética , Drosophila melanogaster/metabolismo , Redes Reguladoras de Genes , MicroARNs/metabolismo , Proteínas Nucleares/metabolismo , Factores de Transcripción/metabolismo , Animales , Femenino , Variación Genética , Genoma de los Insectos , Masculino
2.
Proc Natl Acad Sci U S A ; 117(3): 1312-1320, 2020 01 21.
Artículo en Inglés | MEDLINE | ID: mdl-31900365

RESUMEN

Spherical nucleic acids (SNAs) are nanostructures formed by chemically conjugating short linear strands of oligonucleotides to a nanoparticle template. When made with modified small interfering RNA (siRNA) duplexes, SNAs act as single-entity transfection and gene silencing agents and have been used as lead therapeutic constructs in several disease models. However, the manner in which modified siRNA duplex strands that comprise the SNA lead to gene silencing is not understood. Herein, a systematic analysis of siRNA biochemistry involving SNAs shows that Dicer cleaves the modified siRNA duplex from the surface of the nanoparticle, and the liberated siRNA subsequently functions in a way that is dependent on the canonical RNA interference mechanism. By leveraging this understanding, a class of SNAs was chemically designed which increases the siRNA content by an order of magnitude through covalent attachment of each strand of the duplex. As a consequence of increased nucleic acid content, this nanostructure architecture exhibits less cell cytotoxicity than conventional SNAs without a decrease in siRNA activity.


Asunto(s)
Nanopartículas/química , Interferencia de ARN , ARN Interferente Pequeño/química , Animales , Línea Celular Tumoral , Drosophila melanogaster , Humanos , Nanopartículas/metabolismo , Nanopartículas/toxicidad , ARN Interferente Pequeño/metabolismo , Ribonucleasa III/metabolismo
3.
Appl Environ Microbiol ; 81(3): 1013-23, 2015 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-25416773

RESUMEN

The formation of biofilms is an important survival strategy allowing rhizobia to live on soil particles and plant roots. Within the microcolonies of the biofilm developed by Rhizobium leguminosarum, rhizobial cells interact tightly through lateral and polar connections, forming organized and compact cell aggregates. These microcolonies are embedded in a biofilm matrix, whose main component is the acidic exopolysaccharide (EPS). Our work shows that the O-chain core region of the R. leguminosarum lipopolysaccharide (LPS) (which stretches out of the cell surface) strongly influences bacterial adhesive properties and cell-cell cohesion. Mutants defective in the O chain or O-chain core moiety developed premature microcolonies in which lateral bacterial contacts were greatly reduced. Furthermore, cell-cell interactions within the microcolonies of the LPS mutants were mediated mostly through their poles, resulting in a biofilm with an altered three-dimensional structure and increased thickness. In addition, on the root epidermis and on root hairs, O-antigen core-defective strains showed altered biofilm patterns with the typical microcolony compaction impaired. Taken together, these results indicate that the surface-exposed moiety of the LPS is crucial for proper cell-to-cell interactions and for the formation of robust biofilms on different surfaces.


Asunto(s)
Biopelículas/crecimiento & desarrollo , Lipopolisacáridos/metabolismo , Antígenos O/metabolismo , Raíces de Plantas/microbiología , Rhizobium leguminosarum/fisiología , Lipopolisacáridos/genética , Datos de Secuencia Molecular , Antígenos O/genética , Rhizobium leguminosarum/genética , Rhizobium leguminosarum/crecimiento & desarrollo , Rhizobium leguminosarum/metabolismo , Análisis de Secuencia de ADN
4.
Proc Natl Acad Sci U S A ; 109(30): 12135-40, 2012 Jul 24.
Artículo en Inglés | MEDLINE | ID: mdl-22773814

RESUMEN

Rhizobium leguminosarum is a soil bacterium that infects root hairs and induces the formation of nitrogen-fixing nodules on leguminous plants. Light, oxygen, and voltage (LOV)-domain proteins are blue-light receptors found in higher plants and many algae, fungi, and bacteria. The genome of R. leguminosarum bv. viciae 3841, a pea-nodulating endosymbiont, encodes a sensor histidine kinase containing a LOV domain at the N-terminal end (R-LOV-HK). R-LOV-HK has a typical LOV domain absorption spectrum with broad bands in the blue and UV-A regions and shows a truncated photocycle. Here we show that the R-LOV-HK protein regulates attachment to an abiotic surface and production of flagellar proteins and exopolysaccharide in response to light. Also, illumination of bacterial cultures before inoculation of pea roots increases the number of nodules per plant and the number of intranodular bacteroids. The effects of light on nodulation are dependent on a functional lov gene. The results presented in this work suggest that light, sensed by R-LOV-HK, is an important environmental factor that controls adaptive responses and the symbiotic efficiency of R. leguminosarum.


Asunto(s)
Adhesión Bacteriana/fisiología , Luz , Fotorreceptores Microbianos/metabolismo , Pisum sativum/microbiología , Nodulación de la Raíz de la Planta/fisiología , Rhizobium leguminosarum/fisiología , Simbiosis , Secuencia de Aminoácidos , Adhesión Bacteriana/efectos de la radiación , Secuencia de Bases , Biopelículas/crecimiento & desarrollo , Western Blotting , Flagelos/metabolismo , Violeta de Genciana , Histidina Quinasa , Microscopía Electrónica de Rastreo , Datos de Secuencia Molecular , Nodulación de la Raíz de la Planta/efectos de la radiación , Polisacáridos Bacterianos/metabolismo , Proteínas Quinasas/genética , Proteínas Quinasas/metabolismo , Estructura Terciaria de Proteína/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Rhizobium leguminosarum/efectos de la radiación , Rhizobium leguminosarum/ultraestructura , Alineación de Secuencia , Análisis de Secuencia de ADN , Estadísticas no Paramétricas
5.
J Bacteriol ; 195(20): 4611-9, 2013 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-23935051

RESUMEN

Rhizobia are symbiotic bacteria able to invade and colonize the roots of legume plants, inducing the formation of nodules, where bacteria reduce atmospheric nitrogen (N2) to ammonia (NH3). Riboflavin availability influences the capacity of rhizobia to survive in the rhizosphere and to colonize roots. In this study, we identified the RL1692 gene of Rhizobium leguminosarum downstream of a flavin mononucleotide (FMN) riboswitch. RL1692 encodes a putative transmembrane permease with two EamA domains. The presence of an FMN riboswitch regulating a transmembrane protein is usually observed in riboflavin transporters, suggesting that RL1692 may be involved in riboflavin uptake. The product of RL1692, which we named RibN, is conserved in members of the alpha-, beta-, and gammaproteobacteria and shares no significant identity with any riboflavin transporter previously identified. In this work, we show that RibN is localized in the membrane cellular fraction and its expression is downregulated by riboflavin. By heterologous expression in a Brucella abortus mutant auxotrophic for riboflavin, we demonstrate that RibN possesses flavin transport activity. Similarly, we also demonstrate that RibN orthologues from Ochrobactrum anthropi and Vibrio cholerae (which lacks the FMN riboswitch) are able to transport riboflavin. An R. leguminosarum ribN null mutant exhibited lower nodule occupancy levels in pea plants during symbiosis assays. Thus, we propose that RibN and its homologues belong to a novel family of riboflavin transporters. This work provides the first experimental description of riboflavin transporters in Gram-negative bacteria.


Asunto(s)
Proteínas Bacterianas/metabolismo , Proteínas Portadoras/metabolismo , Regulación Bacteriana de la Expresión Génica/fisiología , Rhizobium leguminosarum/metabolismo , Riboflavina/metabolismo , Secuencia de Aminoácidos , Proteínas Bacterianas/genética , Proteínas Portadoras/genética , Filogenia , Rhizobium leguminosarum/genética
6.
Infect Immun ; 81(3): 996-1007, 2013 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-23319562

RESUMEN

Brucella is responsible for brucellosis, one of the most common zoonoses worldwide that causes important economic losses in several countries. Increasing evidence indicates that adhesion of Brucella spp. to host cells is an important step to establish infection. We have previously shown that the BmaC unipolar monomeric autotransporter mediates the binding of Brucella suis to host cells through cell-associated fibronectin. Our genome analysis shows that the B. suis genome encodes several additional potential adhesins. In this work, we characterized a predicted trimeric autotransporter that we named BtaE. By expressing btaE in a nonadherent Escherichia coli strain and by phenotypic characterization of a B. suis ΔbtaE mutant, we showed that BtaE is involved in the binding of B. suis to hyaluronic acid. The B. suis ΔbtaE mutant exhibited a reduction in the adhesion to HeLa and A549 epithelial cells compared with the wild-type strain, and it was outcompeted by the wild-type strain in the binding to HeLa cells. The knockout btaE mutant showed an attenuated phenotype in the mouse model, indicating that BtaE is required for full virulence. BtaE was immunodetected on the bacterial surface at one cell pole. Using old and new pole markers, we observed that both the BmaC and BtaE adhesins are consistently associated with the new cell pole, suggesting that, in Brucella, the new pole is functionally differentiated for adhesion. This is consistent with the inherent polarization of this bacterium, and its role in the invasion process.


Asunto(s)
Adhesinas Bacterianas/metabolismo , Brucella suis/metabolismo , Brucella suis/patogenicidad , Brucelosis/microbiología , Proteínas Portadoras/metabolismo , Regulación Bacteriana de la Expresión Génica/fisiología , Adhesinas Bacterianas/genética , Animales , Anticuerpos Antibacterianos , Adhesión Bacteriana/fisiología , Brucella suis/genética , Proteínas Portadoras/genética , Polaridad Celular , Escherichia coli/genética , Escherichia coli/metabolismo , Ratones , Ratones Endogámicos BALB C , Familia de Multigenes , Virulencia
7.
Cell Microbiol ; 14(6): 965-82, 2012 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-22321605

RESUMEN

Brucella is an intracellular pathogen responsible of a zoonotic disease called brucellosis. Brucella survives and proliferates within several types of phagocytic and non-phagocytic cells. Like in other pathogens, adhesion of brucellae to host surfaces was proposed to be an important step in the infection process. Indeed, Brucella has the capacity to bind to culture human cells and key components of the extracellular matrix, such as fibronectin. However, little is known about the molecular bases of Brucella adherence. In an attempt to identify bacterial genes encoding adhesins, a phage display library of Brucella suis was panned against fibronectin. Three fibronectin-binding proteins of B. suis were identified using this approach. One of the candidates, designated BmaC was a very large protein of 340 kDa that is predicted to belong to the type I (monomeric) autotransporter family. Microscopy studies showed that BmaC is located at one pole on the bacterial surface. The phage displaying the fibronectin-binding peptide of BmaC inhibited the attachment of brucellae to both, HeLa cells and immobilized fibronectin in vitro. In addition, a bmaC deletion mutant was impaired in the ability of B. suis to attach to immobilized fibronectin and to the surface of HeLa and A549 cells and was out-competed by the wild-type strain in co-infection experiments. Finally, anti-fibronectin or anti-BmaC antibodies significantly inhibited the binding of wild-type bacteria to HeLa cells. Our results highlight the role of a novel monomeric autotransporter protein in the adhesion of B. suis to the extracellular matrix and non-phagocytic cells via fibronectin binding.


Asunto(s)
Adhesinas Bacterianas/fisiología , Adhesión Bacteriana , Brucella suis/fisiología , Interacciones Huésped-Patógeno , Proteínas de Transporte de Membrana/fisiología , Adhesinas Bacterianas/genética , Adhesinas Bacterianas/metabolismo , Animales , Brucella suis/crecimiento & desarrollo , Brucella suis/metabolismo , Fibronectinas/química , Fibronectinas/metabolismo , Técnicas de Inactivación de Genes , Células HeLa , Humanos , Proteínas Inmovilizadas/química , Macrófagos/microbiología , Proteínas de Transporte de Membrana/genética , Proteínas de Transporte de Membrana/metabolismo , Ratones , Viabilidad Microbiana , Biblioteca de Péptidos , Estructura Terciaria de Proteína , Análisis de Secuencia de ADN
8.
Sci Rep ; 13(1): 21172, 2023 12 01.
Artículo en Inglés | MEDLINE | ID: mdl-38040905

RESUMEN

Several COVID-19 vaccines, some more efficacious than others, are now available and deployed, including multiple mRNA- and viral vector-based vaccines. With the focus on creating cost-effective solutions that can reach the low- and medium- income world, GreenLight Biosciences has developed an mRNA vaccine candidate, GLB-COV2-043, encoding for the full-length SARS-CoV-2 Wuhan wild-type spike protein. In pre-clinical studies in mice, GLB-COV2-043 induced robust antigen-specific binding and virus-neutralizing antibody responses targeting homologous and heterologous SARS-CoV-2 variants and a TH1-biased immune response. Boosting mice with monovalent or bivalent mRNA-LNPs provided rapid recall and long-lasting neutralizing antibody titers, an increase in antibody avidity and breadth that was held over time and generation of antigen-specific memory B- and T- cells. In hamsters, vaccination with GLB-COV2-043 led to lower viral loads, reduced incidence of SARS-CoV-2-related microscopic findings in lungs, and protection against weight loss after heterologous challenge with Omicron BA.1 live virus. Altogether, these data indicate that GLB-COV2-043 mRNA-LNP vaccine candidate elicits robust protective humoral and cellular immune responses and establishes our mRNA-LNP platform for subsequent clinical evaluations.


Asunto(s)
COVID-19 , Cricetinae , Animales , Humanos , Ratones , COVID-19/prevención & control , Vacunas contra la COVID-19 , SARS-CoV-2/genética , Modelos Animales , ARN Mensajero/genética , Anticuerpos Neutralizantes , Anticuerpos Antivirales , Inmunogenicidad Vacunal
9.
Front Cell Infect Microbiol ; 11: 607610, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-33987105

RESUMEN

Adhesion to host cells is a key step for successful infection of many bacterial pathogens and may define tropism to different host tissues. To do so, bacteria display adhesins on their surfaces. Brucella is an intracellular pathogen capable of proliferating in a wide variety of cell types. It has been described that BmaC, a large protein that belongs to the classical (type Va) autotransporter family, is required for efficient adhesion of Brucella suis strain 1330 to epithelial cells and fibronectin. Here we show that B. suis 1330 harbors two other type Va autotransporters (BmaA and BmaB), which, although much smaller, share significant sequence similarities with BmaC and contain the essential domains to mediate proper protein translocation to the bacterial surface. Gain and loss of function studies indicated that BmaA, BmaB, and BmaC contribute, to a greater or lesser degree, to adhesion of B. suis 1330 to different cells such as synovial fibroblasts, osteoblasts, trophoblasts, and polarized epithelial cells as well as to extracellular matrix components. It was previously shown that BmaC localizes to a single bacterial pole. Interestingly, we observed here that, similar to BmaC, the BmaB adhesin is localized mostly at a single cell pole, reinforcing the hypothesis that Brucella displays an adhesive pole. Although Brucella species have strikingly similar genomes, they clearly differ in their host preferences. Mainly, the differences identified between species appear to be at loci encoding surface proteins. A careful in silico analysis of the putative type Va autotransporter orthologues from several Brucella strains showed that the bmaB locus from Brucella abortus and both, the bmaA and bmaC loci from Brucella melitensis are pseudogenes in all strains analyzed. Results reported here evidence that all three autotransporters play a role in the adhesion properties of B. suis 1330. However, Brucella spp. exhibit extensive variations in the repertoire of functional adhesins of the classical autotransporter family that can be displayed on the bacterial surface, making them an interesting target for future studies on host preference and tropism.


Asunto(s)
Brucella suis , Sistemas de Secreción Tipo V , Adhesinas Bacterianas/genética , Adhesivos , Brucella abortus , Brucella suis/genética , Sistemas de Secreción Tipo V/genética
10.
Elife ; 92020 02 26.
Artículo en Inglés | MEDLINE | ID: mdl-32101167

RESUMEN

Sensory neuron numbers and positions are precisely organized to accurately map environmental signals in the brain. This precision emerges from biochemical processes within and between cells that are inherently stochastic. We investigated impact of stochastic gene expression on pattern formation, focusing on senseless (sens), a key determinant of sensory fate in Drosophila. Perturbing microRNA regulation or genomic location of sens produced distinct noise signatures. Noise was greatly enhanced when both sens alleles were present in homologous loci such that each allele was regulated in trans by the other allele. This led to disordered patterning. In contrast, loss of microRNA repression of sens increased protein abundance but not sensory pattern disorder. This suggests that gene expression stochasticity is a critical feature that must be constrained during development to allow rapid yet accurate cell fate resolution.


Asunto(s)
Regulación de la Expresión Génica/fisiología , Células Receptoras Sensoriales/metabolismo , Alelos , Animales , Proteínas de Drosophila/metabolismo , Proteínas de Drosophila/fisiología , Drosophila melanogaster/crecimiento & desarrollo , Drosophila melanogaster/metabolismo , Drosophila melanogaster/fisiología , Femenino , Proteínas Nucleares/metabolismo , Proteínas Nucleares/fisiología , Células Receptoras Sensoriales/fisiología , Procesos Estocásticos , Factores de Transcripción/metabolismo , Factores de Transcripción/fisiología , Transcripción Genética
11.
J Bacteriol ; 191(8): 2530-40, 2009 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-19201794

RESUMEN

The RND-type efflux pumps are responsible for the multidrug resistance phenotype observed in many clinically relevant species. Also, RND pumps have been implicated in physiological processes, with roles in the virulence mechanisms of several pathogenic bacteria. We have previously shown that the BepC outer membrane factor of Brucella suis is involved in the efflux of diverse drugs, probably as part of a tripartite complex with an inner membrane translocase. In the present work, we characterize two membrane fusion protein-RND translocases of B. suis encoded by the bepDE and bepFG loci. MIC assays showed that the B. suis DeltabepE mutant was more sensitive to deoxycholate (DOC), ethidium bromide, and crystal violet. Furthermore, multicopy bepDE increased resistance to DOC and crystal violet and also to other drugs, including ampicillin, norfloxacin, ciprofloxacin, tetracycline, and doxycycline. In contrast to the DeltabepE mutant, the resistance profile of B. suis remained unaltered when the other RND gene (bepG) was deleted. However, the DeltabepE DeltabepG double mutant showed a more severe phenotype than the DeltabepE mutant, indicating that BepFG also contributes to drug resistance. An open reading frame (bepR) coding for a putative regulatory protein of the TetR family was found upstream of the bepDE locus. BepR strongly repressed the activity of the bepDE promoter, but DOC released the repression mediated by BepR. A clear induction of the bepFG promoter activity was observed only in the BepDE-defective mutant, indicating a regulatory interplay between the two RND efflux pumps. Although only the BepFG-defective mutant showed a moderate attenuation in model cells, the activities of both bepDE and bepFG promoters were induced in the intracellular environment of HeLa cells. Our results show that B. suis harbors two functional RND efflux pumps that may contribute to virulence.


Asunto(s)
Proteínas Bacterianas/metabolismo , Brucella suis/efectos de los fármacos , Farmacorresistencia Bacteriana , Proteínas de Transporte de Membrana/metabolismo , Secuencia de Aminoácidos , Antibacterianos/farmacología , Proteínas Bacterianas/genética , Brucella suis/patogenicidad , Brucella suis/fisiología , Ácido Desoxicólico/farmacología , Células Epiteliales/microbiología , Etidio/farmacología , Eliminación de Gen , Dosificación de Gen , Regulación Bacteriana de la Expresión Génica , Violeta de Genciana/farmacología , Células HeLa , Humanos , Proteínas de Transporte de Membrana/genética , Pruebas de Sensibilidad Microbiana , Proteínas Represoras/genética , Proteínas Represoras/metabolismo , Alineación de Secuencia , Virulencia
12.
Curr Opin Genet Dev ; 27: 1-6, 2014 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-24791686

RESUMEN

Robustness is a fundamental property of biological systems. The type of robustness that ensures uniform phenotypic outcomes in the face of variation during an organism's development is called canalization. Here, we discuss the roles that microRNAs play in providing canalization to animal development, citing recent theoretical and experimental advances. MicroRNAs repress protein expression, and they do this in ways that create thresholds in expression and provide adaptation to regulatory networks. Numerous examples have now been described where the developmental impact of environmental variation is suppressed by individual microRNAs. A recent paper has found that the impact of genomic variation between individuals is similarly suppressed by a microRNA operating in a developmental network. Thus, genetic variability is held in check, which is potentially important for both animal evolution and manifestation of disease.


Asunto(s)
MicroARNs/genética , Animales , Regulación de la Expresión Génica , Variación Genética , Genoma , Genómica , Humanos , Proteoma/genética
13.
PLoS One ; 8(11): e79770, 2013.
Artículo en Inglés | MEDLINE | ID: mdl-24236157

RESUMEN

The adhesion of bacterial pathogens to host cells is an event that determines infection, and ultimately invasion and intracellular multiplication. Several evidences have recently shown that this rule is also truth for the intracellular pathogen Brucella. Brucella suis displays the unipolar BmaC and BtaE adhesins, which belong to the monomeric and trimeric autotransporter (TA) families, respectively. It was previously shown that these adhesins are involved in bacterial adhesion to host cells and components of the extracellular matrix (ECM). In this work we describe the role of a new member of the TA family of B. suis (named BtaF) in the adhesive properties of the bacterial surface. BtaF conferred the bacteria that carried it a promiscuous adhesiveness to various ECM components and the ability to attach to an abiotic surface. Furthermore, BtaF was found to participate in bacterial adhesion to epithelial cells and was required for full virulence in mice. Similar to BmaC and BtaE, the BtaF adhesin was expressed in a small subpopulation of bacteria, and in all cases, it was detected at the new pole generated after cell division. Interestingly, BtaF was also implicated in the resistance of B. suis to porcine serum. Our findings emphasize the impact of TAs in the Brucella lifecycle.


Asunto(s)
Adhesinas Bacterianas/metabolismo , Adhesión Bacteriana/fisiología , Brucella suis/fisiología , Brucella suis/patogenicidad , Adhesinas Bacterianas/química , Adhesinas Bacterianas/inmunología , Animales , Brucelosis/inmunología , Brucelosis/metabolismo , Línea Celular , Matriz Extracelular/metabolismo , Humanos , Masculino , Ratones , Familia de Multigenes , Multimerización de Proteína , Transporte de Proteínas , Porcinos , Virulencia
14.
Infect Immun ; 75(1): 379-89, 2007 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-17088356

RESUMEN

Brucella spp., like other pathogens, must cope with the environment of diverse host niches during the infection process. In doing this, pathogens evolved different type of transport systems to help them survive and disseminate within the host. Members of the TolC family have been shown to be involved in the export of chemically diverse molecules ranging from large protein toxins to small toxic compounds. The role of proteins from the TolC family in Brucella and other alpha-2-proteobacteria has been explored little. The gene encoding the unique member of the TolC family from Brucella suis (BepC) was cloned and expressed in an Escherichia coli mutant disrupted in the gene encoding TolC, which has the peculiarity of being involved in diverse transport functions. BepC fully complemented the resistance to drugs such as chloramphenicol and acriflavine but was incapable of restoring hemolysin secretion in the tolC mutant of E. coli. An insertional mutation in the bepC gene strongly affected the resistance phenotype of B. suis to bile salts and toxic chemicals such as ethidium bromide and rhodamine and significantly decreased the resistance to antibiotics such as erythromycin, ampicillin, tetracycline, and norfloxacin. Moreover, the B. suis bepC mutant was attenuated in the mouse model of infection. Taken together, these results suggest that BepC-dependent efflux processes of toxic compounds contribute to B. suis survival inside the host.


Asunto(s)
Antiinfecciosos/farmacología , Proteínas de la Membrana Bacteriana Externa/genética , Proteínas de la Membrana Bacteriana Externa/metabolismo , Brucella suis/efectos de los fármacos , Brucella suis/patogenicidad , Resistencia a Medicamentos/genética , Animales , Clonación Molecular , Femenino , Proteínas de Transporte de Membrana/genética , Proteínas de Transporte de Membrana/metabolismo , Ratones , Ratones Endogámicos BALB C , Filogenia , Reacción en Cadena de la Polimerasa , Virulencia
15.
J Bacteriol ; 188(12): 4474-86, 2006 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-16740954

RESUMEN

The type I protein secretion system of Rhizobium leguminosarum bv. viciae encoded by the prsD and prsE genes is responsible for secretion of the exopolysaccharide (EPS)-glycanases PlyA and PlyB. The formation of a ring of biofilm on the surface of the glass in shaken cultures by both the prsD and prsE secretion mutants was greatly affected. Confocal laser scanning microscopy analysis of green-fluorescent-protein-labeled bacteria showed that during growth in minimal medium, R. leguminosarum wild type developed microcolonies, which progress to a characteristic three-dimensional biofilm structure. However, the prsD and prsE secretion mutants were able to form only an immature biofilm structure. A mutant disrupted in the EPS-glycanase plyB gene showed altered timing of biofilm formation, and its structure was atypical. A mutation in an essential gene for EPS synthesis (pssA) or deletion of several other pss genes involved in EPS synthesis completely abolished the ability of R. leguminosarum to develop a biofilm. Extracellular complementation studies of mixed bacterial cultures confirmed the role of the EPS and the modulation of the biofilm structure by the PrsD-PrsE secreted proteins. Protein analysis identified several additional proteins secreted by the PrsD-PrsE secretion system, and N-terminal sequencing revealed peptides homologous to the N termini of proteins from the Rap family (Rhizobium adhering proteins), which could have roles in cellular adhesion in R. leguminosarum. We propose a model for R. leguminosarum in which synthesis of the EPS leads the formation of a biofilm and several PrsD-PrsE secreted proteins are involved in different aspects of biofilm maturation, such as modulation of the EPS length or mediating attachment between bacteria.


Asunto(s)
Transportadoras de Casetes de Unión a ATP/fisiología , Proteínas Bacterianas/metabolismo , Biopelículas/crecimiento & desarrollo , Polisacáridos Bacterianos/fisiología , Rhizobium leguminosarum/fisiología , Ácidos , Proteínas Bacterianas/fisiología , Glicósido Hidrolasas , Polisacáridos Bacterianos/metabolismo
SELECCIÓN DE REFERENCIAS
DETALLE DE LA BÚSQUEDA