18 research outputs found

    The role of the anaphase-promoting complex/cyclosome in plant growth

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    The anaphase-promoting complex/cyclosome (APC/C) is a multi-subunit E3 ubiquitin ligase that plays a major role in the progression of the eukaryotic cell cycle. This unusual protein complex targets key cell cycle regulators, such as mitotic cyclins and securins, for degradation via the 26S proteasome by ubiquitination, triggering the metaphase-to-anaphase transition and exit from mitosis. Because of its essential role in cell cycle regulation, the APC/C has been extensively studied in mammals and yeasts, but relatively less in plants. Evidence shows that, besides its well-known role in cell cycle regulation, the APC/C also has functions beyond the cell cycle. In metazoans, the APC/C has been implicated in cell differentiation, disease control, basic metabolism and neuronal survival. Recent studies also have shed light on specific functions of the APC/C during plant development. Plant APC/C subunits and activators have been reported to play a role in cellular differentiation, vascular development, shoot branching, female and male gametophyte development and embryogenesis. Here, we discuss our current understanding of the APC/C controlling plant growth

    AIP1 is a novel Agenet/Tudor domain protein from Arabidopsis that interacts with regulators of DNA replication, transcription and chromatin remodeling

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    Background: DNA replication and transcription are dynamic processes regulating plant development that are dependent on the chromatin accessibility. Proteins belonging to the Agenet/Tudor domain family are known as histone modification "readers" and classified as chromatin remodeling proteins. Histone modifications and chromatin remodeling have profound effects on gene expression as well as on DNA replication, but how these processes are integrated has not been completely elucidated. It is clear that members of the Agenet/Tudor family are important regulators of development playing roles not well known in plants. Methods: Bioinformatics and phylogenetic analyses of the Agenet/Tudor Family domain in the plant kingdom were carried out with sequences from available complete genomes databases. 3D structure predictions of Agenet/Tudor domains were calculated by I-TASSER server. Protein interactions were tested in two-hybrid, GST pulldown, semi-in vivo pulldown and Tandem Affinity Purification assays. Gene function was studied in a T-DNA insertion GABI-line. Results: In the present work we analyzed the family of Agenet/Tudor domain proteins in the plant kingdom and we mapped the organization of this family throughout plant evolution. Furthermore, we characterized a member from Arabidopsis thaliana named AIP1 that harbors Agenet/Tudor and DUF724 domains. AIP1 interacts with ABAP1, a plant regulator of DNA replication licensing and gene transcription, with a plant histone modification "reader" (LHP1) and with non modified histones. AIP1 is expressed in reproductive tissues and its down-regulation delays flower development timing. Also, expression of ABAP1 and LHP1 target genes were repressed in flower buds of plants with reduced levels of AIP1. Conclusions: AIP1 is a novel Agenet/Tudor domain protein in plants that could act as a link between DNA replication, transcription and chromatin remodeling during flower development

    Silencing CHALCONE SYNTHASE in maize impedes the incorporation of tricin into lignin and increases lignin content

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    Lignin is a phenolic heteropolymer that is deposited in secondary-thickened cell walls, where it provides mechanical strength. A recent structural characterization of cell walls from monocot species showed that the flavone tricin is part of the native lignin polymer, where it is hypothesized to initiate lignin chains. In this study, we investigated the consequences of altered tricin levels on lignin structure and cell wall recalcitrance by phenolic profiling, nuclear magnetic resonance, and saccharification assays of the naturally silenced maize (Zea mays) C2-Idf (inhibitor diffuse) mutant, defective in the CHALCONE SYNTHASE Colorless2 (C2) gene. We show that the C2-Idf mutant produces highly reduced levels of apigenin-and tricin-related flavonoids, resulting in a strongly reduced incorporation of tricin into the lignin polymer. Moreover, the lignin was enriched in beta-beta and beta-5 units, lending support to the contention that tricin acts to initiate lignin chains and that, in the absence of tricin, more monolignol dimerization reactions occur. In addition, the C2-Idf mutation resulted in strikingly higher Klason lignin levels in the leaves. As a consequence, the leaves of C2-Idf mutants had significantly reduced saccharification efficiencies compared with those of control plants. These findings are instructive for lignin engineering strategies to improve biomass processing and biochemical production

    Combining growth-promoting genes leads to positive epistasis in Arabidopsis thaliana

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    Several genes have been described to positively influence final leaf size in Arabidopsis when mutated or overexpressed. However, the connections between these growth regulating genes are still poorly understood. Clearly such knowledge would significantly contribute to understand the biological processes driving leaf growth. In this study, we performed a combinatorial screen with thirteen transgenic Arabidopsis lines with an increased leaf size. Surprisingly, we found that from 61 analyzed combinations, 39% showed an additional increase in leaf size and most of these resulted from a positive epistasis on growth. Similar to what is found in other organisms in which such an epistasis assay was performed, only few genes were highly connected in synergistic combinations. We also observed a positive epistasis in the majority of the combinations with samba, BR11(OE) or SAUR19(OE), suggesting that these growth regulators are more prone to lead to synergistic effects in binary combinations. Furthermore, positive epistasis was not only found with combinations of genes with a similar mode of action, but also with genes which affect distinct processes, such as cell proliferation and cell expansion

    Caracterización ultrasonográfica modo-B y Doppler del cuerpo lúteo en llamas

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    Mode B and Doppler ultrasonography in ruminants, as a technique, has allowed establishing new concepts on the reproductive physiology of females, through the study of follicular dynamics and morphometry of the corpus lu-teum, therefore, the objective was to characterize using mode B ultrasound and Doppler the corpus luteum in recipients’ llamas. 37 llamas were used as recipients (27 for mode B and 10 Doppler ultrasound) that were syn-chronized by applying buserelin acetate 0.0096 mg, nine days later 0.048 mg of Prostaglandin F2α analog was applied, two days later the second dose of buserelin acetate in the same dose to a ovulation and subsequent for-mation of the corpus luteum, The B-mode evaluation of the morphometry consisted of observing the ecotexture, area, diameter and volume of the corpus luteum with the use of a SonoStar SS-8® ultrasound machine at 6.5 MHz frequency and 6 cm depth equipped with a transrectal linear transducer, with the same technique with a Draminski 4Vet® Doppler device, the percentage of luteal area of vascularization (% AVL) was determined. The data were subjected to a descriptive analysis, being 0.119±0.032 cm2 in area, 12.7±1.7 mm in diameter and 0.84±0.32 cm3 in volumen of CL, to determine the relationship between metric measurements the Pearson correlation was used observing a positive relationship between area/diameter of 0.6602, observing a high positive correlation between area and volume, finally, the characteristics of % AVL was 34.97 for the CLs of recipients llamas. In conclusion, the morphometric characteristics evaluated in mode B (area, diameter, and volume) have a positive correlation and the % AVL could be applied as a tool in efficient reproduc-tive management for the selection of receptors in camelids, understanding that it is one of the first reports of these characteristics in llamas. 0.7506, area/volume of 0.9289 and diameter/volume of La ultrasonografía modo B y Doppler en los rumiantes, como técnica ha permitido establecer nuevos conceptos sobre la fisiología reproductiva de las hembras, a través del estudio de la dinámica folicular y morfometría del cuerpo lúteo, por tanto, el objetivo fue caracterizar mediante ultrasonografía modo B y Doppler el cuerpo lúteo en llamas receptoras. Se utilizaron 37 llamas como receptoras (27 para ultrasonografía modo B y 10 Doppler) que fueron sincronizadas aplicando acetato de buserelina 0.0096 mg, nueve días posteriores se aplicó 0.048 mg de análogo de prostaglandina F2α, a los dos días posteriores se aplicó la segunda dosis de acetato de buserelina en la misma dosis para garantizar la ovulación y posterior formación del cuerpo lúteo, la evaluación en modo B de la morfometría consistió en observar la ecotextura, área, diámetro y volumen del cuerpo lúteo con el uso de un equipo ultrasonógrafo SonoStar SS-8® a 6.5 MHz de frecuencia y 6 cm de profundidad equipado con un transductor lineal transrectal; con la misma técnica con un equipo Draminski 4Vet® Doppler se determinó el porcentaje de área luteal de vascularización (% AVL). Los datos fueron sometidos a un análisis descriptivo siendo de 0.119±0.032 cm2 de área, 12.7±1.7 mm de diámetro y 0.84±0.32 cm3 de volumen de CL, para determinar la relación entre medidas métricas se utilizó la correlación de Pearson observando una relación positiva entre área/diámetro de 0.7506, área/volumen de 0.9289 y diámetro/volumen de 0.6602, observando una correlación alta positiva entre área y volumen, finalmente las características de % AVL fue de 34.97 para los CL de llamas receptoras. En conclusión, las características morfométricas evaluadas en modo B (área, diámetro y volumen) tienen una correlación positiva y el % AVL podría aplicarse como herramienta en el manejo reproductivo eficiente para la selección de receptoras en camélidos, entendiendo que es uno de los primeros reportes de estas características en llamas

    Impact of treatment and clinical characteristics on the survival of children with medulloblastoma in Mexico

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    IntroductionData on medulloblastoma outcomes and experiences in low- and middle-income countries, especially in Latin America, is limited. This study examines challenges in Mexico’s healthcare system, focusing on assessing outcomes for children with medulloblastoma in a tertiary care setting.MethodsA retrospective analysis was conducted, involving 284 patients treated at 21 pediatric oncology centers in Mexico.ResultsHigh-risk patients exhibited markedly lower event-free survival than standard-risk patients (43.5% vs. 78.3%, p<0.001). Influential factors on survival included anaplastic subtype (HR 2.4, p=0.003), metastatic disease (HR 1.9, p=0.001); residual tumor >1.5cm², and lower radiotherapy doses significantly impacted event-free survival (EFS) and overall survival (OS). Platinum-based chemotherapy showed better results compared to the ICE protocol in terms of OS and EFS, which was associated with higher toxicity. Patients under 3 years old displayed notably lower OS and EFS compared to older children (36.1% vs. 55.9%, p=0.01)

    The Arabidopsis anaphase promoting complex (APC): regulation through subunit availability in plant tissues

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    Sister-chromatid separation and exit from mitosis require ubiquitin-mediated proteolysis of cell cycle regulators such as cyclin B and securin. The specificity of the reaction is controlled by an ubiquitin-ligase multiprotein complex known as APC (Anaphase Promoting Complex). Comparison of the coding sequences of Arabidopsis genes with the Genbank database reveals extensive homology of the predicted ORFs with the corresponding proteins of other eukaryotes, indicating that the APC is well conserved in plants. However, different from other eukaryotes, the Arabidopsis genes have some particular characteristics, such as the presence of two copies of the CDC27 gene. Furthermore, expression analyses of the AtAPC genes disclose complex profiles that differ, depending on the tissue examined. In actively dividing cell suspensions there is a direct correspondence between the rates of proliferation and mRNA levels from the AtAPC components. On the other hand, in plant organs, dark-grown seedlings and during leaf growth, this correlation is lost and the AtAPC genes are highly expressed in tissues with low overall cell division. Moreover, expression patterns diverge between the subunit genes, raising the possibility that there could be more than one form of the APC, which would execute distinct functions during plant development. The results suggest that an important layer of regulation of APC/C in plants could operate through subunit availability in specific tissues and/or cellular compartments

    Overexpression of the Arabidopsis anaphase promoting complex subunit CDC27a increases growth rate and organ size

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    The Anaphase Promoting Complex (APC) controls CDK activity by targeting the ubiquitin-dependent proteolysis of S-phase and mitosis-promoting cyclins. Here, we report that the ectopic expression of the Arabidopsis CDC27a, an APC subunit, accelerates plant growth and results in plants with increased biomass production. CDC27a overexpression was associated to apical meristem restructuration, protoplasts with higher H-3-thimidine incorporation and altered cell-cycle marker expression. Total protein extracts immunoprecipitated with a CDC27a antibody showed ubiquitin ligase activity, indicating that the Arabidopsis CDC27a gets incorporated into APC complexes. These results indicate a role of AtCDC27a in regulation of plant growth and raise the possibility that the activity of the APC and the rates of plant cell division could be regulated by the concentration of the CDC27a subunit
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