Your browser doesn't support javascript.
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 87.240
Filtrar
1.
J Agric Food Chem ; 68(10): 3267-3276, 2020 Mar 11.
Artículo en Inglés | MEDLINE | ID: mdl-32101430

RESUMEN

Cross-talk between various hormones is important in regulating many aspects of plant growth, development, and senescence, including fruit ripening. Here, exogenous ethylene (ETH, 100 µL/L, 12 h) rapidly accelerated 'Hayward' kiwifruit (Actinidia deliciosa) softening and ethylene production and was enhanced by supplementing with continuous treatment with methyl jasmonate (MeJA, 100 µM/L, 12 h) (ETH+MeJA). ETH+MeJA enhanced ACC synthase (ACS) activities and 1-aminocyclopropane-1-carboxylic acid (ACC) accumulation but not ACC oxidase (ACO) activity. Increased transcripts of ACS genes AdACS1 and AdACS2, ACS activity, and ethylene production were positively correlated. The abundance of AdACS1 was about 6-fold higher than AdACS2. RNA-seq identified 6 transcription factors among the 87 differentially expressed unigenes induced by ETH+MeJA. Dual-luciferase and electrophoretic mobility shift assays (EMSA) indicated that AdNAC2/3 physically interacted with and trans-activated the AdACS1 promoter 2.2- and 3.5-fold, respectively. Collectively, our results indicate that MeJA accelerates ethylene production in kiwifruit induced by exogenous ethylene, via a preferential activation of AdACS1 and AdACS2.


Asunto(s)
Acetatos/farmacología , Actinidia/efectos de los fármacos , Coenzima A Ligasas/metabolismo , Ciclopentanos/farmacología , Etilenos/biosíntesis , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Oxilipinas/farmacología , Reguladores del Crecimiento de las Plantas/farmacología , Proteínas de Plantas/metabolismo , Factores de Transcripción/metabolismo , Actinidia/enzimología , Actinidia/genética , Actinidia/metabolismo , Frutas/efectos de los fármacos , Frutas/enzimología , Frutas/genética , Frutas/metabolismo , Proteínas de Plantas/genética , Factores de Transcripción/genética
2.
Plant Mol Biol ; 102(6): 603-614, 2020 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-32052233

RESUMEN

The WRKY transcription factor family is involved in responding to biotic and abiotic stresses. Its members contain a typical WRKY domain and can regulate plant physiological responses by binding to W-boxes in the promoter regions of downstream target genes. We identified the sweet sorghum SbWRKY50 (Sb09g005700) gene, which encodes a typical class II of the WRKY family protein that localizes to the nucleus and has transcriptional activation activity. The expression of SbWRKY50 in sweet sorghum was reduced by salt stress, and its ectopic expression reduced the salt tolerance of Arabidopsis thaliana plants. Compared with the wild type, the germination rate, root length, biomass and potassium ion content of SbWRKY50 over-expression plants decreased significantly under salt-stress conditions, while the hydrogen peroxide, superoxide anion and sodium ion contents increased. Real-time PCR results showed that the expression levels of AtSOS1, AtHKT1 and genes related to osmotic and oxidative stresses in over-expression strains decreased under salt-stress conditions. Luciferase complementation imaging and yeast one-hybrid assays confirmed that SbWRKY50 could directly bind to the upstream promoter of the SOS1 gene in A. thaliana. However, in sweet sorghum, SbWRKY50 could directly bind to the upstream promoters of SOS1 and HKT1. These results suggest that the new WRKY transcription factor SbWRKY50 participates in plant salt response by controlling ion homeostasis. However, the regulatory mechanisms are different in sweet sorghum and Arabidopsis, which may explain their different salt tolerance levels. The data provide information that can be applied to genetically modifying salt tolerance in different crop varieties.


Asunto(s)
Homeostasis , Tolerancia a la Sal/fisiología , Sorghum/genética , Sorghum/metabolismo , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Biomasa , Proteínas Portadoras , Proteínas de Transporte de Catión/genética , Proteínas de Transporte de Catión/metabolismo , Regulación de la Expresión Génica de las Plantas , Germinación , Peróxido de Hidrógeno/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Raíces de Plantas/crecimiento & desarrollo , Plantas Modificadas Genéticamente , Potasio/metabolismo , Regiones Promotoras Genéticas , Especies Reactivas de Oxígeno/metabolismo , Semillas , Sodio/metabolismo , Intercambiadores de Sodio-Hidrógeno/genética , Intercambiadores de Sodio-Hidrógeno/metabolismo , Estrés Fisiológico , Superóxidos/metabolismo , Simportadores/genética , Simportadores/metabolismo
3.
Artículo en Inglés | MEDLINE | ID: mdl-32017989

RESUMEN

Steroid hormones have been proven as a key drive of sex change in sequentially hermaphroditic organisms. However, the upstream mechanism of sex steroid hormones regulation that affect sex change remain unknown. The main glucocorticoid in teleost fish is cortisol, which both regulates steroidogenesis and has antistress action. Thus, cortisol might be one of the prime factors in sex change. In this study, the glucocorticoid-induced leucine zipper (GILZ) gene, was proven to have a dramatic effect in orange-spotted groupers (Epinephelus coioides) during sex change at the early stage of gonadal transition. The specific action of the GILZ protein is at the pouch-shaped proliferative spermatogonia instead of the degenerative oocyte at the onset of sex change. Immunohistochemical (IHC) evidence revealed that GILZ performs intensively at undifferentiated spermatogonia in the early testis stage. These results imply that cortisol provokes a rise of GILZ through regulation caused by steroid hormones leading to sex change.


Asunto(s)
Lubina/metabolismo , Proteínas de Peces/metabolismo , Leucina Zippers/fisiología , Factores de Transcripción/metabolismo , Secuencia de Aminoácidos , Animales , Lubina/genética , Lubina/crecimiento & desarrollo , Femenino , Proteínas de Peces/biosíntesis , Proteínas de Peces/genética , Gónadas/crecimiento & desarrollo , Gónadas/metabolismo , Organismos Hermafroditas , Masculino , Filogenia , Homología de Secuencia de Aminoácido , Diferenciación Sexual/fisiología , Factores de Transcripción/biosíntesis , Factores de Transcripción/genética
4.
Genes Dev ; 34(3-4): 226-238, 2020 02 01.
Artículo en Inglés | MEDLINE | ID: mdl-31919190

RESUMEN

Centromeres are maintained epigenetically by the presence of CENP-A, an evolutionarily conserved histone H3 variant, which directs kinetochore assembly and hence centromere function. To identify factors that promote assembly of CENP-A chromatin, we affinity-selected solubilized fission yeast CENP-ACnp1 chromatin. All subunits of the Ino80 complex were enriched, including the auxiliary subunit Hap2. Chromatin association of Hap2 is Ies4-dependent. In addition to a role in maintenance of CENP-ACnp1 chromatin integrity at endogenous centromeres, Hap2 is required for de novo assembly of CENP-ACnp1 chromatin on naïve centromere DNA and promotes H3 turnover on centromere regions and other loci prone to CENP-ACnp1 deposition. Prior to CENP-ACnp1 chromatin assembly, Hap2 facilitates transcription from centromere DNA. These analyses suggest that Hap2-Ino80 destabilizes H3 nucleosomes on centromere DNA through transcription-coupled histone H3 turnover, driving the replacement of resident H3 nucleosomes with CENP-ACnp1 nucleosomes. These inherent properties define centromere DNA by directing a program that mediates CENP-ACnp1 assembly on appropriate sequences.


Asunto(s)
Cromatina/metabolismo , Proteínas de Schizosaccharomyces pombe/metabolismo , Schizosaccharomyces/genética , Schizosaccharomyces/metabolismo , Transcripción Genética/fisiología , Cromatina/genética , Proteínas Cromosómicas no Histona/metabolismo , Cromosomas Fúngicos/genética , Cromosomas Fúngicos/metabolismo , ADN de Hongos/metabolismo , Proteínas de Schizosaccharomyces pombe/genética , Factores de Transcripción/metabolismo
5.
Gene ; 732: 144368, 2020 Mar 30.
Artículo en Inglés | MEDLINE | ID: mdl-31954859

RESUMEN

The INO80 complex, including the Ino80 protein, forms a highly conserved canonical complex that remodels chromatin in the context of multiple cellular functions. The Drosophila homologue, dIno80, is involved in homeotic gene regulation during development as a canonical Pho-dIno80 complex. Previously, we found that dIno80 regulates homeotic genes by interacting with epigenetic regulators, such as polycomb and trithorax, suggesting the occurrence of non-canonical Ino80 complexes. Here using spectroscopic methods and gel retardation assays, we identified a set of consensus DNA sequences that DNA binding domain of dIno80 (DBINO) interacts with having differential affinity and high specificity. Testing these sequences in reporter assays, showed that this interaction can positively regulate transcription. These results suggest that, dIno80 has a sequence preference for interaction with DNA leading to transcriptional changes.


Asunto(s)
Proteínas de Unión al ADN/fisiología , ADN/metabolismo , Proteínas de Drosophila/fisiología , Factores de Transcripción/fisiología , Animales , Núcleo Celular/metabolismo , Proteínas de Unión al ADN/metabolismo , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/genética , Drosophila melanogaster/metabolismo , Regulación de la Expresión Génica , Genes Reporteros , Factores de Transcripción/metabolismo , Transcripción Genética
6.
Plant Mol Biol ; 102(6): 615-624, 2020 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-31997111

RESUMEN

KEY MESSAGE: PTR2 in Arabidopsis thaliana is negatively regulated by ABI4 and plays a key role in water uptake by seeds, ensuring that imbibed seeds proceed to germination. Peptide transporters (PTRs) transport nitrogen-containing substrates in a proton-dependent manner. Among the six PTRs in Arabidopsis thaliana, the physiological role of the tonoplast-localized, seed embryo abundant PTR2 is unknown. In the present study, a molecular physiological analysis of PTR2 was conducted using ptr2 mutants and PTR2CO complementation lines. Compared with the wild type, the ptr2 mutant showed ca. 6 h delay in testa rupture and consequently endosperm rupture because of 17% lower water content and 10% higher free abscisic acid (ABA) content. Constitutive overexpression of the PTR2 gene under the control of the Cauliflower mosaic virus (CaMV) 35S promoter in ptr2 mutants rescued the mutant phenotypes. After cold stratification, a transient increase in ABA INSENSITIVE4 (ABI4) transcript levels during induction of testa rupture was followed by a similar increase in PTR2 transcript levels, which peaked prior to endosperm rupture. The PTR2 promoter region containing multiple CCAC motifs was recognized by ABI4 in electrophoretic mobility shift assays, and PTR2 expression was repressed by 67% in ABI4 overexpression lines compared with the wild type, suggesting that PTR2 is an immediate downstream target of ABI4. Taken together, the results suggest that ABI4-dependent temporal regulation of PTR2 expression may influence water status during seed germination to promote the post-germinative growth of imbibed seeds.


Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Transporte Biológico/fisiología , Germinación/fisiología , Proteínas de Transporte de Membrana/metabolismo , Semillas/metabolismo , Agua/metabolismo , Ácido Abscísico/metabolismo , Arabidopsis/genética , Arabidopsis/crecimiento & desarrollo , Proteínas de Arabidopsis/genética , Regulación del Desarrollo de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Germinación/genética , Mutación , Fenotipo , Regiones Promotoras Genéticas , Factores de Transcripción/metabolismo
7.
Nat Commun ; 11(1): 583, 2020 Jan 29.
Artículo en Inglés | MEDLINE | ID: mdl-31996670

RESUMEN

Medulloblastoma (MB) is the most common malignant brain tumor in children and among the subtypes, Group 3 MB has the worst outcome. Here, we perform an in vivo, patient-specific screen leading to the identification of Otx2 and c-MYC as strong Group 3 MB inducers. We validated our findings in human cerebellar organoids where Otx2/c-MYC give rise to MB-like organoids harboring a DNA methylation signature that clusters with human Group 3 tumors. Furthermore, we show that SMARCA4 is able to reduce Otx2/c-MYC tumorigenic activity in vivo and in human cerebellar organoids while SMARCA4 T910M, a mutant form found in human MB patients, inhibits the wild-type protein function. Finally, treatment with Tazemetostat, a EZH2-specific inhibitor, reduces Otx2/c-MYC tumorigenesis in ex vivo culture and human cerebellar organoids. In conclusion, human cerebellar organoids can be efficiently used to understand the role of genes found altered in cancer patients and represent a reliable tool for developing personalized therapies.


Asunto(s)
Neoplasias Encefálicas/metabolismo , Neoplasias Encefálicas/patología , Neoplasias Cerebelosas/metabolismo , Neoplasias Cerebelosas/patología , Meduloblastoma/metabolismo , Meduloblastoma/patología , Organoides/metabolismo , Organoides/patología , Benzamidas/antagonistas & inhibidores , Carcinogénesis , Línea Celular Tumoral , Neoplasias Cerebelosas/genética , ADN Helicasas/genética , ADN Helicasas/metabolismo , Metilación de ADN , Proteína Potenciadora del Homólogo Zeste 2/antagonistas & inhibidores , Regulación Neoplásica de la Expresión Génica , Humanos , Meduloblastoma/genética , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Factores de Transcripción Otx/genética , Factores de Transcripción Otx/metabolismo , Proteínas Proto-Oncogénicas c-myc/genética , Proteínas Proto-Oncogénicas c-myc/metabolismo , Piridonas/antagonistas & inhibidores , Células Madre , Factores de Transcripción/genética , Factores de Transcripción/metabolismo
8.
Plant Mol Biol ; 102(3): 339-357, 2020 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-31894455

RESUMEN

KEY MESSAGE: NCP1, a NINJA family protein lacking EAR motif, acts as a negative regulator of ABA signaling by interacting with and inhibiting the activity of transcriptional activator ABP9. The phytohormone abscisic acid plays a pivotal role in regulating plant responses to a variety of abiotic stresses including drought and salinity. Maize ABP9 is an ABRE-binding bZIP transcription activator that enhances plant tolerance to multiple stresses by positively regulating ABA signaling, but the molecular mechanism by which ABP9 is regulated in mediating ABA responses remains unknown. Here, we report the identification of an ABP9-interacting protein, named ABP Nine Complex Protein 1 (NCP1) and its functional characterization. NCP1 belongs to the recently identified NINJA family proteins, but lacks the conserved EAR motif, which is a hallmark of this class of transcriptional repressors. In vitro and in vivo assays confirmed that NCP1 physically interacts with ABP9 and that they are co-localized in the nucleus. In addition, NCP1 and ABP9 are similarly induced with similar patterns by ABA treatment and osmotic stress. Interestingly, NCP1 over-expressing Arabidopsis plants exhibited a reduced sensitivity to ABA and decreased drought tolerance. Transient assay in maize protoplasts showed that NCP1 inhibits the activity of ABP9 in activating ABRE-mediated reporter gene expression, a notion further supported by genetic analysis of drought and ABA responses in the transgenic plants over-expressing both ABP9 and NCP1. These data together suggest that NCP1 is a novel negative regulator of ABA signaling via interacting with and inhibiting the activity of ABP9.


Asunto(s)
Sequías , Reguladores del Crecimiento de las Plantas/farmacología , Proteínas de Plantas/metabolismo , Transducción de Señal/efectos de los fármacos , Factores de Transcripción/metabolismo , Zea mays/metabolismo , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Genes de Plantas , Presión Osmótica , Reguladores del Crecimiento de las Plantas/genética , Proteínas de Plantas/genética , Plantas Modificadas Genéticamente/genética , Plantas Modificadas Genéticamente/metabolismo , Proteínas Represoras/metabolismo , Salinidad , Estrés Fisiológico , Factores de Transcripción/genética , Transcriptoma , Zea mays/genética
9.
J Agric Food Chem ; 68(3): 918-926, 2020 Jan 22.
Artículo en Inglés | MEDLINE | ID: mdl-31899636

RESUMEN

Theanine (thea) is the most abundant free amino acid in tea plant (Camellia sinensis) and one of the most important secondary metabolites conferring tea quality and health benefits. Great effort has recently been made to functionally dissect enzyme genes (e.g., GS, GDH, GOGAT) responsible for in vivo thea accumulation. However, the transcriptional regulation of its biosynthesis remains to be explored. Starting from publicly available (condition-independent) tea transcriptome data, we performed an exhaustive coexpression analysis between transcription factor (TF) genes and thea enzyme genes in tea plant. Our results showed that two typical plant-specialized (secondary) metabolites related TF families, such as MYB, bHLH, together with WD40 domain proteins, were prominently involved, suggesting a potential MYB-bHLH-WD40 (MBW) complex-mediated regulatory pattern in thea pathway. Aiming at the most involved MYB family, we screened seven MYB genes as thea candidate regulators through a stringent multistep selection (e.g., filtering with condition-specific nitrogen-treated transcriptome data). The control of MYB regulators in thea biosynthesis was further demonstrated using an integrated analysis of thea accumulation and MYB expression in several major tea tissues, including leave, bud, root, and stem. Our investigation aided tea researchers in having a comprehensive view of transcriptional regulatory landscape in thea biosynthesis, serving as the first platform for studying molecular regulation in thea pathway and a paradigm for understanding the characteristic components biosynthesis in nonmodel plants.


Asunto(s)
Camellia sinensis/genética , Camellia sinensis/metabolismo , Glutamatos/biosíntesis , Factores de Transcripción/metabolismo , Vías Biosintéticas , Regulación de la Expresión Génica de las Plantas , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Factores de Transcripción/genética , Transcriptoma
10.
J Agric Food Chem ; 68(5): 1286-1296, 2020 Feb 05.
Artículo en Inglés | MEDLINE | ID: mdl-31891496

RESUMEN

Carotenoids are a class of bioactive compounds that exhibit health-promoting properties for humans, but their regulation in bananas during fruit ripening remains largely unclear. Here, we found that the total carotenoid content continued to be elevated along the course of banana ripening and peaked at the ripening stage followed by a decrease, which is presumably caused by the transcript abundances of carotenoid biosynthetic genes MaLCYB1.1 and MaLCYB1.2. Moreover, a ripening-inducible transcription factor MaSPL16 was characterized, which was a nuclear protein with transactivation activity. Transient transformation of MaSPL16 in banana fruits led to enhanced transcript levels of MaLCYB1.1 and MaLCYB1.2 and hence the total carotenoid accumulation. Importantly, MaSPL16 stimulated the transcription of MaLCYB1.1 and MaLCYB1.2 through directly binding to their promoters. Collectively, our findings indicate that MaSPL16 behaves as an activator to modulate banana carotenoid biosynthesis, which may provide a new target for molecular improvement of the nutritional and bioactive qualities of agricultural crops that accumulate carotenoids.


Asunto(s)
Carotenoides/metabolismo , Frutas/crecimiento & desarrollo , Liasas Intramoleculares/genética , Musa/metabolismo , Proteínas de Plantas/genética , Factores de Transcripción/metabolismo , Frutas/enzimología , Frutas/genética , Frutas/metabolismo , Regulación de la Expresión Génica de las Plantas , Liasas Intramoleculares/metabolismo , Musa/enzimología , Musa/genética , Musa/crecimiento & desarrollo , Proteínas de Plantas/metabolismo , Factores de Transcripción/genética
11.
Nat Commun ; 11(1): 540, 2020 Jan 28.
Artículo en Inglés | MEDLINE | ID: mdl-31992709

RESUMEN

Eukaryotic transcription factors (TF) function by binding to short 6-10 bp DNA recognition sites located near their target genes, which are scattered through vast genomes. Such process surmounts enormous specificity, efficiency and celerity challenges using a molecular mechanism that remains poorly understood. Combining biophysical experiments, theory and bioinformatics, we dissect the interplay between the DNA-binding domain of Engrailed, a Drosophila TF, and the regulatory regions of its target genes. We find that Engrailed binding affinity is strongly amplified by the DNA regions flanking the recognition site, which contain long tracts of degenerate recognition-site repeats. Such DNA organization operates as an antenna that attracts TF molecules in a promiscuous exchange among myriads of intermediate affinity binding sites. The antenna ensures a local TF supply, enables gene tracking and fine control of the target site's basal occupancy. This mechanism illuminates puzzling gene expression data and suggests novel engineering strategies to control gene expression.


Asunto(s)
Sitios de Unión , ADN/metabolismo , Proteínas de Drosophila/metabolismo , Eucariontes/genética , Regulación de la Expresión Génica , Proteínas de Homeodominio/metabolismo , Factores de Transcripción/metabolismo , Animales , Secuencia de Bases , Fenómenos Biofísicos , Biología Computacional , Proteínas de Unión al ADN , Drosophila melanogaster/genética , Drosophila melanogaster/metabolismo , Genoma , Cinética , Modelos Teóricos , Conformación Molecular , Termodinámica
12.
Nat Commun ; 11(1): 74, 2020 01 03.
Artículo en Inglés | MEDLINE | ID: mdl-31900393

RESUMEN

Despite the promising clinical efficacy of the second-generation anaplastic lymphoma kinase (ALK) inhibitor alectinib in patients with ALK-rearranged lung cancer, some tumor cells survive and eventually relapse, which may be an obstacle to achieving a cure. Limited information is currently available on the mechanisms underlying the initial survival of tumor cells against alectinib. Using patient-derived cell line models, we herein demonstrate that cancer cells survive a treatment with alectinib by activating Yes-associated protein 1 (YAP1), which mediates the expression of the anti-apoptosis factors Mcl-1 and Bcl-xL, and combinatorial inhibition against both YAP1 and ALK provides a longer tumor remission in ALK-rearranged xenografts when compared with alectinib monotherapy. These results suggest that the inhibition of YAP1 is a candidate for combinatorial therapy with ALK inhibitors to achieve complete remission in patients with ALK-rearranged lung cancer.


Asunto(s)
Proteínas Adaptadoras Transductoras de Señales/metabolismo , Quinasa de Linfoma Anaplásico/genética , Apoptosis/efectos de los fármacos , Carbazoles/administración & dosificación , Reordenamiento Génico/efectos de los fármacos , Neoplasias Pulmonares/tratamiento farmacológico , Recurrencia Local de Neoplasia/tratamiento farmacológico , Piperidinas/administración & dosificación , Factores de Transcripción/metabolismo , Proteínas Adaptadoras Transductoras de Señales/genética , Quinasa de Linfoma Anaplásico/metabolismo , Animales , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Femenino , Humanos , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/fisiopatología , Masculino , Ratones , Ratones Endogámicos BALB C , Recurrencia Local de Neoplasia/genética , Recurrencia Local de Neoplasia/metabolismo , Recurrencia Local de Neoplasia/fisiopatología , Inhibidores de Proteínas Quinasas/administración & dosificación , Factores de Transcripción/genética
13.
BMC Bioinformatics ; 21(1): 3, 2020 Jan 02.
Artículo en Inglés | MEDLINE | ID: mdl-31898480

RESUMEN

BACKGROUND: Observed levels of gene expression strongly depend on both activity of DNA binding transcription factors (TFs) and chromatin state through different histone modifications (HMs). In order to recover the functional relationship between local chromatin state, TF binding and observed levels of gene expression, regression methods have proven to be useful tools. They have been successfully applied to predict mRNA levels from genome-wide experimental data and they provide insight into context-dependent gene regulatory mechanisms. However, heterogeneity arising from gene-set specific regulatory interactions is often overlooked. RESULTS: We show that regression models that predict gene expression by using experimentally derived ChIP-seq profiles of TFs can be significantly improved by mixture modelling. In order to find biologically relevant gene clusters, we employ a Bayesian allocation procedure which allows us to integrate additional biological information such as three-dimensional nuclear organization of chromosomes and gene function. The data integration procedure involves transforming the additional data into gene similarity values. We propose a generic similarity measure that is especially suitable for situations where the additional data are of both continuous and discrete type, and compare its performance with similar measures in the context of mixture modelling. CONCLUSIONS: We applied the proposed method on a data from mouse embryonic stem cells (ESC). We find that including additional data results in mixture components that exhibit biologically meaningful gene clusters, and provides valuable insight into the heterogeneity of the regulatory interactions.


Asunto(s)
Células Madre Embrionarias/metabolismo , Regulación de la Expresión Génica , Células Madre Pluripotentes/metabolismo , Animales , Teorema de Bayes , Cromatina/genética , Cromatina/metabolismo , Inmunoprecipitación de Cromatina , Genoma , Ratones , Análisis de Regresión , Factores de Transcripción/genética , Factores de Transcripción/metabolismo
14.
Dev Genes Evol ; 230(1): 39-45, 2020 01.
Artículo en Inglés | MEDLINE | ID: mdl-31960123

RESUMEN

The molluscan larval shell formation is a complicated process. There is evidence that the mantle of the primary larva (trochophore) contains functionally different cell populations with distinct gene expression profiles. However, it remains unclear how these cells are specified. In the present study, we identified three cell populations from the shell gland in earlier stages (gastrula) from the bivalve mollusc Crassostrea gigas. These cell populations were determined by analyzing the co-expression relationships among six potential shell formation (pSF) genes using two-color hybridization. The three cell populations, which we designated as SGCPs (shell gland cell populations), formed a concentric-circle pattern from outside to inside of the shell gland. SGCP I was located in the outer edge of the shell gland and the cells expressed pax2/5/8, gata2/3, and bmp2/4. SGCP II was located more internally and the cells expressed two engrailed genes. The last population, SGCP III, was located in the central region of the shell gland and the cells expressed lox4. Determination of the gene expression profiles of SGCPs would help trace their origins and fates and elucidate how these cell populations are specified. Moreover, potential roles of the SGCPs, e.g., development of sensory cells and shell biogenesis, are suggested. Our results reveal the internal organization of the embryonic shell gland at the molecular level and add to the knowledge of larval shell formation.


Asunto(s)
Crassostrea/citología , Exoesqueleto/citología , Exoesqueleto/metabolismo , Animales , Crassostrea/genética , Crassostrea/crecimiento & desarrollo , Crassostrea/metabolismo , Glándulas Exocrinas/citología , Glándulas Exocrinas/metabolismo , Femenino , Masculino , Factores de Transcripción/metabolismo
15.
Adv Exp Med Biol ; 1217: 173-186, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-31898228

RESUMEN

Posttranslational protein modifications play an important role in regulating protein stability and cellular function. There are at least eight Cullin family members. Among them, Cullin-2 forms a functional E3 ligase complex with elongin B, elongin C, RING-box protein 1 (RBX1, also called ROC1), as well as the substrate recognition subunit (SRS) to promote the substrate ubiquitination and degradation. In this book chapter, we will review Cullin-2 E3 ligase complexes that include various SRS proteins, including von Hippel Lindau (pVHL), leucine-rich repeat protein-1 (LRR-1), preferentially expressed antigen of melanoma (PRAME), sex-determining protein FEM-1 and early embryogenesis protein ZYG-11. We will focus on the VHL signaling pathway in clear cell renal cell carcinoma (ccRCC), which may reveal various therapeutic avenues in treating this lethal cancer.


Asunto(s)
Carcinoma de Células Renales/enzimología , Proteínas Cullin/metabolismo , Neoplasias Renales/enzimología , Animales , Carcinoma de Células Renales/tratamiento farmacológico , Humanos , Neoplasias Renales/tratamiento farmacológico , Unión Proteica , Factores de Transcripción/metabolismo , Proteína Supresora de Tumores del Síndrome de Von Hippel-Lindau/metabolismo
16.
Adv Exp Med Biol ; 1217: 187-210, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-31898229

RESUMEN

Cullin 3 (Cul3) family of ubiquitin ligases comprises three components, the RING finger protein RBX1, the Cul3 scaffold, and a Bric-a-brac/Tramtrack/Broad complex (BTB) protein. The BTB protein serves as a bridge to connect Cul3 to substrate and is functionally equivalent to the combination of substrate adaptor and linker in other Cullin complexes. Human genome encodes for ~180 BTB proteins, implying a broad spectrum of ubiquitination signals and substrate repertoire. Accordingly, Cul3 ubiquitin ligases are involved in diverse cellular processes, including cell division, differentiation, cytoskeleton remodeling, stress responses, and nerve cell functions. Emerging evidence has pointed to the prominent role of Cul3 ubiquitin ligases in cancer. This chapter will describe recent advances on the roles of Cul3 E3 ligase complexes in regulating various cancer hallmarks and therapeutic responses and the mutation/dysregulation of Cul3 substrate adaptors in cancer. In particular, we will focus on several extensively studied substrate adaptors, such as Keap1, SPOP, KLHL20, and LZTR1, and will also discuss other recently identified Cul3 adaptors with oncogenic or tumor-suppressive functions. We conclude that Cul3 ubiquitin ligases represent master regulators of human malignancies and highlight the importance of developing modulating agents for oncogenic/tumor-suppressive Cul3 E3 ligase complexes to prevent or intervene tumorigenesis.


Asunto(s)
Carcinogénesis , Proteínas Cullin/metabolismo , Neoplasias/metabolismo , Neoplasias/patología , Proteínas Adaptadoras Transductoras de Señales/metabolismo , Animales , Humanos , Proteína 1 Asociada A ECH Tipo Kelch/metabolismo , Proteínas Nucleares/metabolismo , Proteínas Represoras/metabolismo , Factores de Transcripción/metabolismo
17.
Cell Prolif ; 53(2): e12757, 2020 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-31916327

RESUMEN

OBJECTIVES: To testify that endothelial cells (ECs) induce astrocyte maturation by leukaemia inhibitory factor (LIF) secretion. MATERIALS AND METHODS: In vivo experiments, mice bearing floxed alleles of YAP were crossed with mice expressing a Cre recombinase driven by the endothelial Tek promoter (Tek-Cre) to finally obtain the following three genotypes: YAPf/f , Tek-Cre; YAPf/w , Tek-Cre; and YAPf/f . Retinal vascularization and astrocyte network were evaluated by whole-mount fluorescence and Western blotting. In vitro, experiments were performed in an astrocyte and human microvascular endothelial cell (HMEC-1) coculture model to analyse the mechanisms underlying the effect of endothelial YAP on astrocytes. RESULTS: In vivo, YAPf/f ;Tek-Cre mice showed delayed angiogenesis, sparse vessels and decreased glial fibrillary acidic protein (GFAP)+ astrocytes but aberrant growth of endothelial networks and immature astrocytes (platelet-derived growth factor A, PDGFRA+ astrocytes) overgrowth. In vitro, Yap deletion attenuated the LIF release that delayed the maturation of retinal astrocyte which was consistent with the results of HMEC-1-astrocyte coculture. The effect of YAP overexpression on LIF-LIFR axis in HMEC-1 interferes the GFAP expression of astrocyte. In contrast, LIF protein rescues the astrocytic GFAP expression when EC YAP was inhibited by siRNAs. CONCLUSIONS: We show that EC yes-associated protein (YAP) is not only a critical coactivator of Hippo signalling in retinal vessel development but also plays an essential role in retinal astrocyte maturation by regulating LIF production.


Asunto(s)
Proteínas Adaptadoras Transductoras de Señales/metabolismo , Astrocitos/metabolismo , Factor Inhibidor de Leucemia/metabolismo , Retina/metabolismo , Vasos Retinianos/metabolismo , Factores de Transcripción/metabolismo , Animales , Astrocitos/fisiología , Diferenciación Celular/fisiología , Proliferación Celular/fisiología , Técnicas de Cocultivo/métodos , Células Endoteliales/metabolismo , Células Endoteliales/fisiología , Femenino , Proteína Ácida Fibrilar de la Glía/metabolismo , Humanos , Masculino , Ratones , Ratones Endogámicos C57BL , Neovascularización Patológica/metabolismo , Neovascularización Patológica/patología , Neovascularización Fisiológica/fisiología , Neurogénesis/fisiología , Retina/fisiología , Vasos Retinianos/fisiología
18.
mSphere ; 5(1)2020 Jan 29.
Artículo en Inglés | MEDLINE | ID: mdl-31996420

RESUMEN

Toxoplasma gondii can infect and replicate in vascular endothelial cells prior to entering host tissues. However, little is known about the molecular interactions at the parasite-endothelial cell interface. We demonstrate that T. gondii infection of primary human umbilical vein endothelial cells (HUVEC) altered cell morphology and dysregulated barrier function, increasing permeability to low-molecular-weight polymers. T. gondii disrupted vascular endothelial cadherin (VE-cadherin) and ß-catenin localization to the cell periphery and reduced VE-cadherin protein expression. Notably, T. gondii infection led to reorganization of the host cytoskeleton by reducing filamentous actin (F-actin) stress fiber abundance under static and microfluidic shear stress conditions and by reducing planar cell polarity. RNA sequencing (RNA-Seq) comparing genome-wide transcriptional profiles of infected to uninfected endothelial cells revealed changes in gene expression associated with cell-cell adhesion, extracellular matrix reorganization, and cytokine-mediated signaling. In particular, genes downstream of Hippo signaling and the biomechanical sensor and transcriptional coactivator Yes-associated protein (YAP) were downregulated in infected endothelial cells. Interestingly, T. gondii infection activated Hippo signaling by increasing phosphorylation of LATS1, leading to cytoplasmic retention of YAP, and reducing YAP target gene expression. These findings suggest that T. gondii infection triggers Hippo signaling and YAP nuclear export, leading to an altered transcriptional profile of infected endothelial cells.IMPORTANCE Toxoplasma gondii is a foodborne parasite that infects virtually all warm-blooded animals and can cause severe disease in individuals with compromised or weakened immune systems. During dissemination in its infected hosts, T. gondii breaches endothelial barriers to enter tissues and establish the chronic infections underlying the most severe manifestations of toxoplasmosis. The research presented here examines how T. gondii infection of primary human endothelial cells induces changes in cell morphology, barrier function, gene expression, and mechanotransduction signaling under static conditions and under the physiological conditions of shear stress found in the bloodstream. Understanding the molecular interactions occurring at the interface between endothelial cells and T. gondii may provide insights into processes linked to parasite dissemination and pathogenesis.


Asunto(s)
Permeabilidad de la Membrana Celular , Células Endoteliales de la Vena Umbilical Humana/parasitología , Mecanotransducción Celular , Toxoplasma/patogenicidad , Actinas/metabolismo , Proteínas Adaptadoras Transductoras de Señales/metabolismo , Antígenos CD/metabolismo , Cadherinas/metabolismo , Polaridad Celular , Células Cultivadas , Citoesqueleto , Células Endoteliales de la Vena Umbilical Humana/citología , Humanos , Proteínas Serina-Treonina Quinasas/metabolismo , Fibras de Estrés/metabolismo , Factores de Transcripción/metabolismo , Transcriptoma , beta Catenina/metabolismo
19.
J Agric Food Chem ; 68(2): 530-540, 2020 Jan 15.
Artículo en Inglés | MEDLINE | ID: mdl-31891490

RESUMEN

The influence of ß-hydroxy-ß-methylbutyrate (HMB) on proliferation and differentiation of myogenic cells has been well-studied. However, the role of HMB in myofiber specification and potential mechanisms is largely unknown. Thus, the objective of this research was to explore the role of HMB supplementation in myofiber specification. Results showed that HMB treatment significantly increased the fast MyHC protein level (mice: 1.59 ± 0.08, P < 0.01; C2C12: 2.26 ± 0.11, P < 0.001), decreased the slow MyHC protein level (mice: 0.76 ± 0.05, P < 0.05; C2C12: 0.52 ± 0.02, P < 0.001), and increased the miR-199a-3p level (mice: 4.93 ± 0.37, P < 0.001; C2C12: 11.25 ± 0.57, P < 0.001). Besides, we also observed that HMB promoted the activity of glycolysis-related enzymes and reduced the activities of oxidation-related enzymes in mice and C2C12 cells. Overexpression of miR-199a-3p downregulated the slow MyHC protein level (0.71 ± 0.02, P < 0.01) and upregulated the fast MyHC protein level (2.13 ± 0.09, P < 0.001), while repression of miR-199a-3p exhibited the opposite effect. Target identification results verified that miR-199a-3p targets the 3'UTR of the TEA domain family member 1 (TEAD1) to cause its post-transcriptional inhibition (0.41 ± 0.07, P < 0.01). Knockdown of TEAD1 exhibited a similar effect with miR-199a-3p on myofiber specification. Moreover, suppression of miR-199a-3p blocked slow-to-fast myofiber type transition induced by HMB. Together, our finding revealed that miR-199-3p is induced by HMB and contributes to the action of HMB on slow-to-fast myofiber type conversion via targeting TEAD1.


Asunto(s)
MicroARNs/metabolismo , Fibras Musculares Esqueléticas/efectos de los fármacos , Mioblastos Esqueléticos/efectos de los fármacos , Valeratos/farmacología , Regiones no Traducidas 3' , Animales , Diferenciación Celular/efectos de los fármacos , Proteínas de Unión al ADN/genética , Proteínas de Unión al ADN/metabolismo , Ratones , MicroARNs/genética , Fibras Musculares Esqueléticas/citología , Fibras Musculares Esqueléticas/metabolismo , Mioblastos Esqueléticos/citología , Mioblastos Esqueléticos/metabolismo , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Regulación hacia Arriba/efectos de los fármacos
20.
Biochim Biophys Acta Rev Cancer ; 1873(1): 188341, 2020 01.
Artículo en Inglés | MEDLINE | ID: mdl-31931113

RESUMEN

Understanding the molecular mechanisms driving resistance to anti-cancer drugs is both a crucial step to define markers of response to therapy and a clinical need in many cancer settings. YAP and TAZ transcriptional cofactors behave as oncogenes in different cancer types. Deregulation of YAP/TAZ expression or alterations in components of the multiple signaling pathways converging on these factors are important mechanisms of resistance to chemotherapy, target therapy and hormone therapy. Moreover, response to immunotherapy may also be affected by YAP/TAZ activities in both tumor and microenvironment cells. For these reasons, various compounds inhibiting YAP/TAZ function by different direct and indirect mechanisms have been proposed as a mean to counter-act drug resistance in cancer. A particularly promising approach may be to simultaneously target both YAP/TAZ expression and their transcriptional activity through BET inhibitors.


Asunto(s)
Proteínas Adaptadoras Transductoras de Señales/metabolismo , Antineoplásicos/uso terapéutico , Resistencia a Antineoplásicos/efectos de los fármacos , Neoplasias/tratamiento farmacológico , Transactivadores/metabolismo , Factores de Transcripción/metabolismo , Microambiente Tumoral/efectos de los fármacos , Proteínas Adaptadoras Transductoras de Señales/genética , Resistencia a Antineoplásicos/genética , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Humanos , Redes y Vías Metabólicas/efectos de los fármacos , Redes y Vías Metabólicas/genética , Neoplasias/genética , Neoplasias/metabolismo , Transducción de Señal/efectos de los fármacos , Transducción de Señal/genética , Transactivadores/genética , Factores de Transcripción/genética , Microambiente Tumoral/genética
SELECCIÓN DE REFERENCIAS
DETALLE DE LA BÚSQUEDA