RESUMEN
Tropical forest root characteristics and resource acquisition strategies are underrepresented in vegetation and global models, hampering the prediction of forest-climate feedbacks for these carbon-rich ecosystems. Lowland tropical forests often have globally unique combinations of high taxonomic and functional biodiversity, rainfall seasonality, and strongly weathered infertile soils, giving rise to distinct patterns in root traits and functions compared with higher latitude ecosystems. We provide a roadmap for integrating recent advances in our understanding of tropical forest belowground function into vegetation models, focusing on water and nutrient acquisition. We offer comparisons of recent advances in empirical and model understanding of root characteristics that represent important functional processes in tropical forests. We focus on: (1) fine-root strategies for soil resource exploration, (2) coupling and trade-offs in fine-root water vs nutrient acquisition, and (3) aboveground-belowground linkages in plant resource acquisition and use. We suggest avenues for representing these extremely diverse plant communities in computationally manageable and ecologically meaningful groups in models for linked aboveground-belowground hydro-nutrient functions. Tropical forests are undergoing warming, shifting rainfall regimes, and exacerbation of soil nutrient scarcity caused by elevated atmospheric CO2. The accurate model representation of tropical forest functions is crucial for understanding the interactions of this biome with the climate.
Las características de las raíces de los bosques tropicales y las estrategias de adquisición de recursos están subrepresentadas en modelos de vegetación, lo que dificulta la predicción del efecto de cambio de clima para estos ecosistemas ricos en carbono. Los bosques tropicales a menudo tienen combinaciones únicas a nivel mundial de alta biodiversidad taxonómica y funcional, estacionalidad de precipitación, y suelos infértiles, dando lugar a patrones distintos en los rasgos y funciones de las raíces en comparación con los ecosistemas de latitudes más altas. Integramos los avances recientes en nuestra comprensión de la función subterránea de los bosques tropicales en modelos de vegetación, centrándonos en la adquisición de agua y nutrientes. Ofrecemos comparaciones de avances recientes en la comprensión empírica y de modelos de las características de las raíces que representan procesos funcionales importantes en los bosques tropicales. Nos centramos en: (1) estrategias de raíces finas para adquisición de recursos del suelo, (2) acoplamiento y compensaciones entre adquisición del agua y de nutrientes, y (3) vínculos entre funciones sobre tierra y debajo del superficie en bosques tropicales. Sugerimos vías para representar estas comunidades de plantas extremadamente diversas en grupos computacionalmente manejables y ecológicamente significativos en modelos. Los bosques tropicales se están calentando, tienen cambios en los regímenes de lluvias, y tienen una exacerbación de la escasez de nutrientes del suelo causada por el elevado CO2 atmosférico. La representación precisa de las funciones de los bosques tropicales en modelos es crucial para comprender las interacciones de este bioma con el clima.
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Ecosistema , Raíces de Plantas , Nitrógeno , Bosques , Suelo , Plantas , Agua , Clima Tropical , ÁrbolesRESUMEN
Tropical ecosystems face escalating global change. These shifts can disrupt tropical forests' carbon (C) balance and impact root dynamics. Since roots perform essential functions such as resource acquisition and tissue protection, root responses can inform about the strategies and vulnerabilities of ecosystems facing present and future global changes. However, root trait dynamics are poorly understood, especially in tropical ecosystems. We analyzed existing research on tropical root responses to key global change drivers: warming, drought, flooding, cyclones, nitrogen (N) deposition, elevated (e) CO2, and fires. Based on tree species- and community-level literature, we obtained 266 root trait observations from 93 studies across 24 tropical countries. We found differences in the proportion of root responsiveness to global change among different global change drivers but not among root categories. In particular, we observed that tropical root systems responded to warming and eCO2 by increasing root biomass in species-scale studies. Drought increased the root: shoot ratio with no change in root biomass, indicating a decline in aboveground biomass. Despite N deposition being the most studied global change driver, it had some of the most variable effects on root characteristics, with few predictable responses. Episodic disturbances such as cyclones, fires, and flooding consistently resulted in a change in root trait expressions, with cyclones and fires increasing root production, potentially due to shifts in plant community and nutrient inputs, while flooding changed plant regulatory metabolisms due to low oxygen conditions. The data available to date clearly show that tropical forest root characteristics and dynamics are responding to global change, although in ways that are not always predictable. This synthesis indicates the need for replicated studies across root characteristics at species and community scales under different global change factors.
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Cambio Climático , Sequías , Raíces de Plantas , Clima Tropical , Raíces de Plantas/crecimiento & desarrollo , Raíces de Plantas/metabolismo , Árboles/crecimiento & desarrollo , Biomasa , Nitrógeno/metabolismo , Bosques , Inundaciones , IncendiosRESUMEN
Mangroves are among the most carbon-dense ecosystems worldwide. Most of the carbon in mangroves is found belowground, and root production might be an important control of carbon accumulation, but has been rarely quantified and understood at the global scale. Here, we determined the global mangrove root production rate and its controls using a systematic review and a recently formalised, spatially explicit mangrove typology framework based on geomorphological settings. We found that global mangrove root production averaged ~770 ± 202 g of dry biomass m-2 year-1 globally, which is much higher than previously reported and close to the root production of the most productive tropical forests. Geomorphological settings exerted marked control over root production together with air temperature and precipitation (r2 ≈ 30%, p < .001). Our review shows that individual global changes (e.g. warming, eutrophication, drought) have antagonist effects on root production, but they have rarely been studied in combination. Based on this newly established root production rate, root-derived carbon might account for most of the total carbon buried in mangroves, and 19 Tg C lost in mangroves each year (e.g. as CO2 ). Inclusion of root production measurements in understudied geomorphological settings (i.e. deltas), regions (Indonesia, South America and Africa) and soil depth (>40 cm), as well as the creation of a mangrove root trait database will push forward our understanding of the global mangrove carbon cycle for now and the future. Overall, this review presents a comprehensive analysis of root production in mangroves, and highlights the central role of root production in the global mangrove carbon budget.
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Carbono , Ecosistema , Humedales , Biomasa , Bosques , SueloRESUMEN
Mangroves are among the world's most carbon-dense ecosystems, but have suffered extensive deforestation, prompting reforestation projects. The effects of mangrove reforestation on belowground carbon dynamics are poorly understood. In particular, we do not know how fine root production develops following mangrove reforestation, despite fine root production being a major carbon sink and an important control of mangrove soil accretion. Using minirhizotrons, we investigated fine root production and its depth variation along a chronosequence of mature Vietnamese mangroves. Our results showed that fine root production decreases strongly with stand age in the uppermost 32 cm of our soil profiles. In younger mangrove stands, fine root production declines with depth, possibly due to a vertical gradient in soil nutrient availability; while root production in the oldest stand is low at all depths and exhibits no clear vertical pattern. A major fraction of fine root production occurs deeper than 30 cm, depths that are commonly omitted from calculations of mangrove carbon budgets. Younger mangroves may accrue shallow soil organic matter faster than older mangroves. Therefore, root productivity and forest stand age should be accounted for when forecasting mangrove carbon budgets and resistance to sea-level rise.
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Ecosistema , Humedales , Carbono , Secuestro de Carbono , Bosques , SueloRESUMEN
Mangroves are among the world's most carbon-dense ecosystems, but they are threatened by rapid climate change and rising sea levels. The accumulation and decomposition of soil organic matter (SOM) are closely tied to mangroves' carbon sink functions and resistance to rising sea levels. However, few studies have investigated the response of mangrove SOM dynamics to likely future environmental conditions. We quantified how mangrove SOM decay is affected by predicted global warming (+4°C), sea level changes (simulated by altering of the inundation duration to 0, 2, and 6 hr/day), and their interaction. Whilst changes in inundation duration between 2 and 6 hr/day did not affect SOM decay, the treatment without inundation led to a 60% increase. A warming of 4°C caused SOM decay to increase by 21%, but longer inundation moderated this temperature-driven increase. Our results indicate that (a) sea level rise is unlikely to decrease the SOM decay rate, suggesting that previous mangrove elevation gain, which has allowed mangroves to persist in areas of sea level rise, might result from changes in root production and/or mineral sedimentation; (b) sea level fall events, predicted to double in frequency and area, will cause periods of intensified SOM decay; (c) changing tidal regimes in mangroves due to sea level rise might attenuate increases in SOM decay caused by global warming. Our results have important implications for forecasting mangrove carbon dynamics and the persistence of mangroves and other coastal wetlands under future scenarios of climate change.
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Ecosistema , Suelo , Secuestro de Carbono , Cambio Climático , HumedalesRESUMEN
CD9, a member of the tetraspanin family, has been implicated in hematopoietic and leukemic stem cell homing. We investigated the role of CD9 in the dissemination of B acute lymphoblastic leukemia (B-ALL) cells, by stably downregulating CD9 in REH and NALM6 cells. CD9 expression was associated with higher levels of REH cell adhesion to fibronectin and C-X-C motif chemokine receptor 4 (CXCR4)-mediated migration. Death occurred later in NOD/SCID mice receiving REH cells depleted of CD9 for transplantation than in mice receiving control cells. After C-X-C motif chemokine ligand 12 (CXCL12) stimulation, CD9 promoted the formation of long cytoplasmic actin-rich protrusions. We demonstrated that CD9 enhanced RAC1 activation, in both REH cells and blasts from patients. Conversely, the overexpression of a competing CD9 C-terminal tail peptide in REH cytoplasm decreased RAC1 activation and cytoplasmic extension formation in response to CXCL12. Finally, the inhibition of RAC1 activation decreased migration in vitro, and the depletion of RAC1 protein from transplanted REH cells increased mouse survival. Furthermore, a testis-conditioned medium induced the migration of REH and NALM6 cells, and this migration was impeded by an anti-CD9 antibody. The level of CD9 expression also influenced the homing of these cells in mouse testes. These findings demonstrate, for the first time, that CD9 plays a key role in the CXCR4-mediated migration and engraftment of B-ALL cells in the bone marrow or testis, through RAC1 activation.
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Movimiento Celular , Regulación Neoplásica de la Expresión Génica , Neuropéptidos/metabolismo , Leucemia-Linfoma Linfoblástico de Células Precursoras B/metabolismo , Leucemia-Linfoma Linfoblástico de Células Precursoras B/patología , Receptores CXCR4/metabolismo , Tetraspanina 29/metabolismo , Proteína de Unión al GTP rac1/metabolismo , Animales , Apoptosis , Western Blotting , Médula Ósea/metabolismo , Médula Ósea/patología , Adhesión Celular , Proliferación Celular , Quimiocina CXCL12/metabolismo , Humanos , Inmunoprecipitación , Masculino , Ratones , Ratones Endogámicos NOD , Ratones SCID , ARN Mensajero/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Transducción de Señal , Testículo/metabolismo , Testículo/patología , Células Tumorales Cultivadas , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
Many argue that monitoring conducted exclusively by scientists is insufficient to address ongoing environmental challenges. One solution entails the use of mobile digital devices in participatory monitoring (PM) programs. But how digital data entry affects programs with varying levels of stakeholder participation, from nonscientists collecting field data to nonscientists administering every step of a monitoring program, remains unclear. We reviewed the successes, in terms of management interventions and sustainability, of 107 monitoring programs described in the literature (hereafter programs) and compared these with case studies from our PM experiences in Australia, Canada, Ethiopia, Ghana, Greenland, and Vietnam (hereafter cases). Our literature review showed that participatory programs were less likely to use digital devices, and 2 of our 3 more participatory cases were also slow to adopt digital data entry. Programs that were participatory and used digital devices were more likely to report management actions, which was consistent with cases in Ethiopia, Greenland, and Australia. Programs engaging volunteers were more frequently reported as ongoing, but those involving digital data entry were less often sustained when data collectors were volunteers. For the Vietnamese and Canadian cases, sustainability was undermined by a mismatch in stakeholder objectives. In the Ghanaian case, complex field protocols diminished monitoring sustainability. Innovative technologies attract interest, but the foundation of effective participatory adaptive monitoring depends more on collaboratively defined questions, objectives, conceptual models, and monitoring approaches. When this foundation is built through effective partnerships, digital data entry can enable the collection of more data of higher quality. Without this foundation, or when implemented ineffectively or unnecessarily, digital data entry can be an additional expense that distracts from core monitoring objectives and undermines project sustainability. The appropriate role of digital data entry in PM likely depends more on the context in which it is used and less on the technology itself.
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Computadores , Conservación de los Recursos Naturales , Recolección de Datos , Monitoreo del Ambiente , Australia , Canadá , Etiopía , Ghana , HumanosRESUMEN
Introduction: antiretroviral therapy enables the suppression of the plasma viral load and the restoration of immune responses. Therapeutic failures are still observed in patients living with HIV despite the considerable benefits of antiretroviral therapy. This study aimed to describe the long-term evolution of immunological and virological parameters in patients undergoing treatments for HIV-1 at the Day Hospital of Bobo-Dioulasso in Burkina Faso. Methods: a retrospective descriptive and analytical study covering 10 years from 2009 was conducted at the Sourô Sanou University Hospital Center (CHUSS) in Bobo-Dioulasso. HIV-1-positive patients with at least two viral load measurements and two CD4 T cell counts were included in this study. Excel 2019 and RStudio were used to analyze the data. Results: a total of 265 patients were included in this study. The mean age of the patients was 48 ± 8.98 years and women accounted for 77.7% of the study population. A considerable decrease in the number of patients with TCD4 lymphocytes below 200 cells/µl from year 2 of treatment and a progressive increase in those with TCD4 lymphocytes above 500 cells/µl were observed in the study. Regarding the evolution of viral load, an increase in the proportions of patients with an undetectable viral load and a decrease in those with a viral load greater than 1000 copies/ml were noticed in years 2, 5, 6, and 8 of the follow-up. However, a decrease in the proportions of patients with undetectable viral load and an increase in those with viral load above 1000 copies/ml were observed in the years 4, 7, and 10 of follow-up. Conclusion: this study highlighted the different trends of viral load and LTCD4 evolution over 10 years of antiretroviral treatment. It showed a good immunovirological response was shown at the beginning of antiretroviral therapy, and then, a poor evolution of these markers at certain periods during the follow-up of HIV-positive patients.
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Infecciones por VIH , VIH-1 , Humanos , Femenino , Adulto , Persona de Mediana Edad , Estudios Retrospectivos , Burkina Faso/epidemiología , Infecciones por VIH/epidemiología , Hospitales UniversitariosRESUMEN
INTRODUCTION: A cross-sectional study by questionnaire was performed to evaluate the knowledge of French dentists about bone resorption inhibitors (BRIs) and medication-related osteonecrosis of the jaw (MRONJ). MATERIALS AND METHODS: 498 digital questionnaires were collected and 358 complete responses of French dental practionners in active practice except for orthodontists were analyzed. Descriptive analysis was computed and categorical variables were compared by Z test. The independence of the compared variables was tested by χ2 test. The scores obtained to the knowledge questions were compared by Mann-Whitney's tests depending on age, gender and year of graduation. RESULTS: 84% of the respondents routinely record antiresorptive medication history in the medical chart. Therefore, almost all the practitioners know the importance to report in anamnesis the use of BRIs, but we noticed some contradictions: Less than half of the respondents recognized the brand names of BRIs and their indications. The combination of BRIs with other drugs like antiangiogenic or corticosteroid therapies is identified as a MRONJ systemic risk factor by respectively 46,3% and 29,7% of the respondents. Likewise, only 43,2% of the practitioners identified removable dentures as a local risk factor. We showed that practitioners under 30 years old and/or who graduated for less than 10 years reached a significantly higher score putting university as the main source of information on that subject. CONCLUSION: According to our results, it is evident that there is a lack of knowledge about BRI, the risk of MRONJ, and the methods and means of preventing this complication.
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Osteonecrosis de los Maxilares Asociada a Difosfonatos , Conservadores de la Densidad Ósea , Adulto , Actitud , Osteonecrosis de los Maxilares Asociada a Difosfonatos/epidemiología , Osteonecrosis de los Maxilares Asociada a Difosfonatos/etiología , Osteonecrosis de los Maxilares Asociada a Difosfonatos/prevención & control , Conservadores de la Densidad Ósea/efectos adversos , Estudios Transversales , Denosumab , Odontólogos , Difosfonatos/efectos adversos , HumanosRESUMEN
Acute lymphoblastic leukemias (ALL) are the most frequent cancer in children and derive most often from B-cell precursors. Current survival rates roughly reach 90% at 10 years from diagnosis. However, 15-20% of children still relapse with a significant risk of death. Our previous work showed that the transmembrane protein CD9 plays a major role in lymphoblasts migration into sanctuary sites, especially in testis, through the activation of RAC1 signaling upon blasts stimulation with C-X-C chemokine ligand 12 (CXCL12). Here, we identified common factors shared by the bone marrow and extramedullary niches which could upregulate CD9 expression and function. We found that low oxygen levels enhance CD9 expression both at mRNA and protein levels. We further determined that Hypoxia Inducible Factor 1α (HIF1α), the master transcription factor involved in hypoxia response, binds directly CD9 promoter and induce CD9 transcription. We also showed that CD9 protein is crucial for leukemic cell adhesion and migration at low oxygen levels, possibly through its action on RAC1 signaling. Mouse xenograft experiments indicate that HIF1α signaling pathway promotes ALL cells engraftment in a CD9-dependent manner. The present work increments our understanding of CD9 implication in ALL pathogenesis.
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Hipoxia , Transducción de Señal , Masculino , Humanos , Ratones , Animales , Tetraspanina 29/genética , Tetraspanina 29/metabolismo , Adhesión Celular , OxígenoRESUMEN
Duchenne muscular dystrophy is the most common and severe form of muscular dystrophy, and although the genetic basis of this disease is well defined, the overall mechanisms that define its pathogenesis remain obscure. Alterations in individual signaling pathways have been described, but little information is available regarding their putative implications in Duchenne muscular dystrophy pathogenesis. Here, we studied the status of various major signaling pathways in the Golden Retriever muscular dystrophy dog that specifically reproduces the full spectrum of human pathology. Using antibody arrays, we found that Akt1, glycogen synthase kinase-3beta (GSK3beta), 70-kDa ribosomal protein S6 kinase (p70S6K), extracellular signal-regulated kinases 1/2, and p38delta and p38gamma kinases all exhibited decreased phosphorylation in muscle from a 4-month-old animal with Golden Retriever muscular dystrophy, revealing a deep alteration of the phosphatidylinositol 3-kinase (PI3K)/Akt and mitogen-activated protein kinase pathways. Immunohistochemistry analysis revealed the presence of muscle fibers exhibiting a cytosolic accumulation of Akt1, GSK3beta, and phosphatidylinositol-3,4,5-trisphosphate 3-phosphatase (PTEN), an enzyme counteracting PI3K-mediated Akt activation. Enzymatic assays established that these alterations in phosphorylation and expression levels were associated with decreased Akt and increased GSK3beta and PTEN activities. PTEN/GSK3beta-positive fibers were also observed in muscle sections from 3- and 36-month-old animals, indicating long-term PI3K/Akt pathway alteration. Collectively, our data suggest that increased PTEN expression and activity play a central role in PI3K/Akt/GSK3beta and p70S6K pathway modulation, which could exacerbate the consequences of dystrophin deficiency.
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Distrofia Muscular Animal/fisiopatología , Fosfohidrolasa PTEN/metabolismo , Fosfatidilinositol 3-Quinasas/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Animales , Western Blotting , Perros , Distrofina/deficiencia , Glucógeno Sintasa Quinasa 3/metabolismo , Glucógeno Sintasa Quinasa 3 beta , Humanos , Inmunohistoquímica , Microscopía Confocal , Músculo Esquelético/metabolismo , Músculo Esquelético/patología , Distrofia Muscular Animal/metabolismo , Distrofia Muscular Animal/patología , Distrofia Muscular de Duchenne/metabolismo , Distrofia Muscular de Duchenne/patología , Distrofia Muscular de Duchenne/fisiopatología , Fosforilación , Proteínas Quinasas S6 Ribosómicas 70-kDa/metabolismo , Transducción de Señal/fisiologíaRESUMEN
BACKGROUND: Fine root production is one of the least well understood components of the carbon cycle in terrestrial ecosystems. Minirhizotrons allow accurate and non-destructive sampling of fine root production. Small and large scale studies across a range of ecosystems are needed to have baseline data on fine root production and further assess the impact of global change upon it; however, the expense and the low adaptability of minirhizotrons prevent such data collection, in worldwide distributed sampling schemes, in low-income countries and in some ecosystems (e.g. tropical forested wetlands). RESULTS: We present EnRoot, a narrow minirhizotron of 25 mm diameter, that is partially 3D printable. EnRoot is inexpensive (150), easy to construct (no prior knowledge required) and adapted to a range of ecosystems including tropical forested wetlands (e.g. mangroves, peatlands). We tested EnRoot's accuracy and precision for measuring fine root length and diameter, and it yielded Lin's concordance correlation coefficient values of 0.95 for root diameter and 0.92 for length. As a proof of concept, we tested EnRoot in a mesocosm study, and in the field in a tropical mangrove. EnRoot proved its capacity to capture the development of roots of a legume (Medicago sativa) and a mangrove species (seedlings of Rhizophora mangle) in laboratory mesocosms. EnRoot's field installation was possible in the root-dense tropical mangrove because its narrow diameter allowed it to be installed between larger roots and because it is fully waterproof. EnRoot compares favourably with commercial minirhizotrons, and can image roots as small as 56 µm. CONCLUSION: EnRoot removes barriers to the extensive use of minirhizotrons by being low-cost, easy to construct and adapted to a wide range of ecosystem. It opens the doors to worldwide distributed minirhizotron studies across an extended range of ecosystems with the potential to fill knowledge gaps surrounding fine root production.
RESUMEN
Serum response factor (SRF) is a MADS transcription factor that binds to the CArG box sequence of the serum response element (SRE). Through its binding to CArG sequences, SRF activates several muscle-specific genes as well as genes that respond to mitogens. The thermodynamic parameters of the interaction of core-SRF (the 124-245 fragment of serum response factor) with specific oligonucleotides from c-fos and desmin promoters, were determined by spectroscopy. The rotational correlation time of core-SRF labeled with bis-ANS showed that the protein is monomeric at low concentration (10(-7) m). The titration curves for the fluorescence anisotropy of fluorescein-labeled oligonucleotide revealed that under equilibrium conditions, the core-SRF monomers were bound sequentially to SRE at very low concentration (10(-9) m). Curve-fitting data showed also major differences between the wild-type sequence and the oligonucleotide sequences mutated within the CArG box. The fluorescence of the core-SRF tyrosines was quenched by the SRE oligonucleotide. This quenching indicated that under stoichiometric conditions, core-SRF was bound as a dimer to the wild-type oligonucleotide, and as a monomer or a tetramer to the mutant oligonucleotides. Far-UV CD spectra indicated that the flexibility of core-SRF changed profoundly upon its binding to its specific target SRE. Lastly, the rotational correlation time of fluorescein-labeled SRE revealed that formation of the specific complex was accompanied by a change in the SRE internal dynamics. These results indicated that the flexibility of the two partners is crucial for the DNA-protein interaction.
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ADN/metabolismo , Modelos Moleculares , Elemento de Respuesta al Suero/fisiología , Factor de Respuesta Sérica/metabolismo , Secuencia de Aminoácidos , Sitios de Unión , Dicroismo Circular , ADN/química , Desmina/genética , Dimerización , Polarización de Fluorescencia , Genes fos , Datos de Secuencia Molecular , Conformación de Ácido Nucleico , Oligonucleótidos/metabolismo , Regiones Promotoras Genéticas , Conformación Proteica , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Factor de Respuesta Sérica/genética , SolucionesRESUMEN
While specific posttranslational modification patterns within the H3 and H4 tail domains are associated with the S-phase, their actual functions in replication-dependent chromatin assembly have not yet been defined. Here we used incorporation of trace amounts of recombinant proteins into naturally synchronous macroplasmodia of Physarum polycephalum to examine the function of H3 and H4 tail domains in replication-coupled chromatin assembly. We found that the H3/H4 complex lacking the H4 tail domain was not efficiently recovered in nuclei, whereas depletion of the H3 tail domain did not impede nuclear import but chromatin assembly failed. Furthermore, our results revealed that the proper pattern of acetylation on the H4 tail domain is required for nuclear import and chromatin assembly. This is most likely due to binding of Hat1, as coimmunoprecipitation experiments showed Hat1 associated with predeposition histones in the cytoplasm and with replicating chromatin. These results suggest that the type B histone acetyltransferase assists in shuttling the H3/H4 complex from cytoplasm to the replication forks.
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Ensamble y Desensamble de Cromatina , Replicación del ADN , Histona Acetiltransferasas/metabolismo , Histonas/metabolismo , Physarum polycephalum/metabolismo , Proteínas Recombinantes de Fusión/metabolismo , Acetilación , Transporte Activo de Núcleo Celular , Histonas/genética , Modelos Biológicos , Estructura Terciaria de Proteína , Proteínas Recombinantes de Fusión/genéticaRESUMEN
Stearoyl-CoA Desaturase-1 (SCD1) is the rate limiting enzyme catalyzing the synthesis of monounsaturated fatty acids. Variation of SCD1 activity and the ratio of saturated to unsaturated fatty acids have been implicated in a variety of diseases including obesity, type II diabetes and cancers. In liver, many factors regulate SCD1 expression including dietary and hormonal factors such as insulin and leptin. We previously showed in hepatic cells that insulin acts through the PI3K and mTOR pathways to upregulate SCD1 expression. In the present study, using HepG2 cells, we characterized the signaling pathway mediating the leptin inhibitory response on SCD1 gene expression. We showed that leptin inhibits SCD1 at the transcriptional level. Inhibition of the ERK1/2 MAPK pathway with the PD98059 reverses the effect of leptin on SCD1 expression. Our data also demonstrated that the effect of leptin is entirely independent of the effect of insulin. Using the pharmaceutical inhibitors Ag490 and SL0101, we showed that the inhibitory effect of leptin is also mediated by the Janus Kinase 2 (Jak2) and p90RSK. EMSA and transfection experiments suggest a key role for the Sp1 transcription factor, which in turn may compete for the binding of other transcription factors such as AP-1, leading to the inhibition of SCD1 transcription. Taken together, our observations showed that, independently of insulin action, leptin exerts an inhibitory effect on SCD1 transcription via a signaling pathway implicating Jak2, ERK1/2, and p90RSK which probably targets the downstream transcription factor Sp1 on the SCD1 promoter.
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Regulación de la Expresión Génica/efectos de los fármacos , Leptina/farmacología , Proteínas Quinasas Activadas por Mitógenos/fisiología , Estearoil-CoA Desaturasa/genética , Transcripción Genética/efectos de los fármacos , Transcripción Genética/fisiología , Análisis de Varianza , Western Blotting , Células Cultivadas , Ensayo de Cambio de Movilidad Electroforética , Células Hep G2 , Humanos , Insulina/farmacología , Janus Quinasa 2/metabolismo , Regiones Promotoras Genéticas/efectos de los fármacos , Regiones Promotoras Genéticas/fisiología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Estearoil-CoA Desaturasa/metabolismoRESUMEN
An array of phage-displayed mimetic peptides representing 18 sequences from the HIV-1 gp41 immunodominant epitope was fabricated on a nitrocellulose membrane. It used to compare the antibody-binding affinity of the peptide and to monitor the immune response of four patients prior to and after the initiation of Highly Active Anti-Retroviral Therapy in parallel assays with several probes using near-infrared fluorescence detection. The proposed multiplexed approach is highly sensitive, consumes less sample and can be used to analyze the immune repertoires of virus-infected individuals.
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Proteína gp41 de Envoltorio del VIH/fisiología , VIH-1/metabolismo , Biblioteca de Péptidos , Proteómica/métodos , Secuencia de Aminoácidos , Anticuerpos/química , Terapia Antirretroviral Altamente Activa , Ensayo de Inmunoadsorción Enzimática , Epítopos/química , Proteína gp41 de Envoltorio del VIH/química , Inmunoglobulina G/química , Datos de Secuencia Molecular , Análisis por Matrices de Proteínas , Unión Proteica , Proteínas Virales/químicaRESUMEN
The protein array methodology is used to study DNA-protein and protein-protein interactions governing gene expression from the Bacillus stearothermophilus PargCo promoter-operator region. Using probes labelled with near-infrared fluorescence dyes with exitation characteristics close to 700 or 800 nm, it is possible to detect signals from proteins (purified or non-purified in Escherichia coli cell extracts) immobilised on a nitrocellulose membrane with a high sensitivity (almost 12 amol of a spotted protein for protein-DNA interactions). Protein array data are confirmed by other methods indicating that molecular interactions of the order 10(-7) M can be monitored with the proposed protein array approach. We show that the PargCo region is a target for binding at least three types of regulatory proteins, ArgR repressors from thermophilic bacteria, the E. coli RNA polymerase alpha subunit and cyclic AMP binding protein CRP. We also demonstrate that the high strength of the PargC promoter is related to an upstream element that binds to the E. coli RNA polymerase alpha subunit.
Asunto(s)
Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , ADN Bacteriano/genética , ADN Bacteriano/metabolismo , Análisis por Matrices de Proteínas/métodos , Secuencia de Bases , Escherichia coli/genética , Escherichia coli/metabolismo , Proteínas de Escherichia coli/genética , Proteínas de Escherichia coli/metabolismo , Genes Bacterianos , Geobacillus stearothermophilus/genética , Geobacillus stearothermophilus/metabolismo , Datos de Secuencia Molecular , Regiones Operadoras Genéticas , Regiones Promotoras Genéticas , Análisis por Matrices de Proteínas/estadística & datos numéricos , Unión Proteica , Proteómica , Sensibilidad y Especificidad , Espectroscopía Infrarroja Corta , Transcripción GenéticaRESUMEN
Bacillus stearothermophilus ArgR binds efficiently to the Escherichia coli carAB operator, whereas the E. coli repressor binds very poorly to the argCo operator of B. stearothermophilus. In order to elucidate this contradictory behavior between ArgRs, we constructed chimeric proteins by swapping N-terminal DNA-binding and C-terminal oligomerization domains or by exchanging the linker peptide. Chimeras carrying the E. coli DNA-binding domain and the B. stearothermophilus oligomerization domain showed sequence-nonspecific rather than sequence-specific interactions with arg operators. Chimeras carrying the B. stearothermophilus DNA-binding domain and E. coli oligomerization domain exhibited a high DNA-binding affinity for the B. stearothermophilus argCo and E. coli carAB operators and repressed the reporter-gene transcription from the B. stearothermophilus PargCo control region in vitro; arginine had no effect on, and indeed even decreased, their DNA-binding affinity. With the protein array method, we showed that the wild-type B. stearothermophilus ArgR and derivatives of it containing only the exchanged linker from E. coli ArgR or carrying the B. stearothermophilus DNA-binding domain along with the linker and the alpha4 regions were able to bind argCo containing the single Arg box. This binding was weaker than binding to the two-box operator but was no longer arginine dependent. Several lines of observations indicate that the alpha4 helix in the oligomerization domain and the linker peptide can contribute to the recognition of single or double Arg boxes and therefore to the operator DNA-binding specificity in similar but not identical ArgR repressors from two distant bacteria.