Deficiencies of essential vitamins, iron (Fe), and zinc (Zn) affect over one-half of the world's population. A significant progress has been made to control micronutrient deficiencies through supplementation, but new approaches are needed, especially to reach the rural poor. Agronomic biofortification of pulses with Zn, Fe, and boron (B) offers a pragmatic solution to combat hidden hunger instead of food fortification and supplementation. Moreover, it also has positive effects on crop production as well. Therefore, we conducted three separate field experiments for two consecutive years to evaluate the impact of soil and foliar application of the aforementioned nutrients on the yield and seed biofortification of mungbean. Soil application of Zn at 0, 4.125, 8.25, Fe at 0, 2.5, 5.0 and B at 0, 0.55, 1.1 kg ha −1 was done in the first, second and third experiment, respectively. Foliar application in these experiments was done at 0.3% Zn, 0.2% Fe and 0.1% B respectively one week after flowering initiation. Data revealed that soil-applied Zn, Fe and B at 8.25, 5.0 and 1.1 kg ha −1 , respectively, enhanced the grain yield of mungbean; however, this increase in yield was statistically similar to that recorded with Zn, Fe and B at 4.125, 2.5 and 0.55 kg ha −1 , respectively. Foliar application of these nutrients at flower initiation significantly enhanced the Zn contents by 28% and 31%, Fe contents by 80% and 78%, while B contents by 98% and 116% over control during 2019 and 2020, respectively. It was concluded from the results that soil application of Zn, Fe, and B enhanced the yield performance of mungbean; while significant improvements in seed Zn, Fe, and B contents were recorded with foliar application of these nutrients.
Allostery plays a key role in dictating the stoichiometry and thermodynamics of multi-protein complexes driving a plethora of cellular processes central to health and disease. Herein, using various biophysical tools, we demonstrate that although Sos1 nucleotide exchange factor and Gab1 docking protein recognize two non-overlapping sites within the Grb2 adaptor, allostery promotes the formation of two distinct pools of Grb2-Sos1 and Grb2-Gab1 binary signaling complexes in concert in lieu of a composite Sos1-Grb2-Gab1 ternary complex. Of particular interest is the observation that the binding of Sos1 to the nSH3 domain within Grb2 sterically blocks the binding of Gab1 to the cSH3 domain and vice versa in a mutually exclusive manner. Importantly, the formation of both the Grb2-Sos1 and Grb2-Gab1 binary complexes is governed by a stoichiometry of 2:1, whereby the respective SH3 domains within Grb2 homodimer bind to Sos1 and Gab1 via multivalent interactions. Collectively, our study sheds new light on the role of allostery in mediating cellular signaling machinery.
Coregulators span a broad and extensive domain in modulating cellular transcriptional activity. Studies have established a dynamic role for such coregulators in various endocrine cancers. Steroid hormone receptors (SHRs) play a pivotal role in such endocrine cancers, and interact abundantly with transcriptional coregulators in altering gene expression. Several families of coregulators have implications in propagating the development, progression and invasion of breast, prostate, and other hormone-responsive cancers. This mini-review aims to discuss different classes of coregulators involved in endocrine cancers and highlight unique information regarding each family with relevance to mechanism, intervention, and novel directions being investigated. V C 2016 IUBMB Life, 68 (7): [504][505][506][507][508][509][510][511][512][513][514][515] 2016
Current study was designed during 2019-2020 at Agricultural Research Farm, Agronomy (Fodder Production) Section, Ayub Agricultural Research Institute, Faisalabad, Pakistan with main objective to evaluate the varying supplementary dietary levels of Moringa oleifera fresh leaves for optimum production in Nili-Ravi buffaloes. A trial of 9 buffaloes was selected having 3-4 parity and were divided into 3 groups: A, B and C. Buffaloes in group A, B and C were supplementary fed 2 kg, 3 kg and 4 kg Moringa oleifera fresh leaves per day for one week each. The influence of Moringa oleifera fresh leaves was studies on following parameters: animal weight gain, milk yield and milk composition. The collected data were analyzed statistically. The result of this study indicated that optimum weight gain was found in the animals supplementarily fed 4 kg Moringa oleifera fresh leaves per day and average body weight of animals increased from 669, 503 and 644 kg to 678, 507 and 651 kg in group A, B and C, respectively. Similarly animals fed 4 kg Moringa oleifera fresh leaves per day yielded optimum milk production as compared to other groups of animals wherein average milk yield was increased from 5.05, 6.50 and 5.45 liters to 5.6, 6.7 and 5.9 liters in group A, B and C, respectively. Total solids in the milk were also increased in the treatment where 4 kg Moringa oleifera fresh leaves were fed to animals per day where fat % increased from 6.5, 6.9 and 6.8 to 6.8, 7.0 and 6.9, Crude protein % from 4.72, 4.5 and 4.51 to 4.81, 4.64 and 4.6 % and calcium % increased from 0.112, 0.117 and 0.101 to 0.13, 0.129 and 0.116 in group A, B and C, respectively. It was concluded that in addition to all other feeding requirements, 4 kg dietary Moringa oleifera fresh leaves might affect the body weight and production performance of Nili-Ravi buffaloes positively without any ill effects on milk aroma and appearance.
Purpose/Objective(s): Steroid hormone receptors, upon hormonal activation, act to modulate gene expression in receptive cells through interactions with coregulator proteins. Protein Kinase A (PKA) is a particularly potent downstream effector, and is conservatively regulated in a number of biochemical pathways. The current project aims to elucidate the mechanisms of action of E6AP, a known critical player in endocrine cancer on the expression/activity of PKA in neuroblastoma. Materials/Methods: A microarray genetic analysis yielded a decreased PKA-R2A expression dependent upon decreased levels of estradiol (E2) or E6AP. This correlation was validated via a PCR determined downregulation of PKA-R2A mRNA with E6AP knockdown in Neuro2a cells. Bioinformatics provided four potential Estrogen Response Element (ERE) sequences in the PKA-R2A promoter region. These sequences were cloned to a Luciferase reporter gene vector. HeLa cells harboring these constructs were incubated with and without E2. Transcriptional activity was surrogated by bioluminescence via a Luciferase-Luciferin reaction, normalized to Renilla luminal output. Electrophoretic mobility shift assay (EMSA) was performed to detect direct ER binding to biotin-labeled ERE in the presence and absence of E2. ER-specific antibodies generated a super-shift. Unlabeled and consensus ERE allowed for competitive binding. Chromatin immunoprecipitation (ChIP) investigated recruitment of the complete transcriptional complex E6AP-ER onto the promoter region of the PKA-R2A gene and its association with active expression of the gene in Neuro2a cells. Results: A Luciferase luminal output to unit Renilla luminal output ratio was obtained in the absence and presence of E2. The scrambled control yielded ratios of 2.77 and 1.97, respectively. The consensus control yielded 9.55 and 85.37. ERE-1 yielded significant ratios of 8.57 and 55.7. EMSA evaluation of ERE-1 demonstrated a biotin-labeled ERE band shift in the presence of ER incubated with E2 relative to that without E2, and a super-shift with addition of an Anti-ER antibody. Competitive saturation with both the unlabeled and consensus ERE sequences negated these observed shifts. ChIP results show, upon E2 treatment in Neuro2a cells, increased recruitment of the E6AP-ER complex to the same ERE-1 associated with increased H3K14 acetylation and p300 recruitment at the same site which was coupled with the recruitment of pRNAPolII to the transcriptional start site of the PKA-R2A gene indicating active transcription and expression of the gene. Conclusion: ERE sequences exist and appear to function in response to E2 within the PKA-R2A promoter region, modulating the transcriptional status of a ubiquitous and crucial protein. Novel therapies targeting this hormonal foundation may potentially play a role in ameliorating the symptomatic manifestations associated with neuroblastoma. Citation Format: Jean-Pierre Obeid, Nawal Zafar, Jimmy El Hokayem. E6-associated protein-dependent estrogen receptor modulation of protein kinase A regulatory subunit R2A expression in neuroblastoma. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1851.
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