Akt-Induced Phosphorylation of N-CoR at Serine 1450 Contributes to Its Misfolded Conformational Dependent Loss (MCDL) in Acute Myeloid Leukemia of the M5 Subtype

The nuclear receptor co-repressor (N-CoR) is a key component of the generic co-repressor complex that plays an important role in the control of cellular growth and differentiation. As shown by us recently, the growth suppressive function of N-CoR largely relies on its capacity to repress Flt3, a key regulator of cellular gorwth during normal and malignant hematopoesis. We further demonstrated how de-repression of Flt3 due to the misfolded conformation dependent loss (MCDL) of N-CoR contributed to malignant growth in acute myeloid leukemia (AML). However, the molecular mechanism underlying the MCDL of N-CoR and its implication in AML pathogenesis is not fully understood. Here, we report that Akt-induced phosphorylation of N-CoR at the consensus Akt motif is crucial for its misfolding and subsequent loss in AML (AML-M5). N-CoR displayed significantly higher level of serine specific phosphorylation in almost all AML-M5 derived cells and was subjected to processing by AML-M5 specific aberrant protease activity. To identify the kinase linked to N-CoR phosphorylation, a library of activated kinases was screened with the extracts of AML cells; leading to the identification of Akt as the putative kinase linked to N-CoR phosphorylation. Consistent with this finding, a constitutively active Akt consistently phosphorylated N-CoR leading to its misfolding; while the therapeutic and genetic ablation of Akt largely abrogated the MCDL of N-CoR in AML-M5 cells. Site directed mutagenic analysis of N-CoR identified serine 1450 as the crucial residue whose phosphorylation by Akt was essential for the misfolding and loss of N-CoR protein. Moreover, Akt-induced phosphorylation of N-CoR contributed to the de-repression of Flt3, suggesting a cross talk between Akt signaling and N-CoR misfolding pathway in the pathogenesis of AML-M5. The N-CoR misfolding pathway could be the common downstream thread of pleiotropic Akt signaling activated by various oncogenic insults in some subtypes of leukemia and solid tumors.     

Accuracy checks of physical beam modifier factors algorithm used in computerized treatment planning system for a 15 MV photon beam

In order to optimize the tumour dose by using wedge filters, systematic studies were carried out to investigate the accuracy of the beam modifier algorithm in a computerized treatment planning system (Theraplan plus, version 3.8). The effect of different parameters such as beam hardening and softening coefficients on the wedge factor was also studied. A 15 MV photon beam obtained from a linear accelerator was used throughout the experiments. Normalized wedge factors were determined experimentally as well as with the Theraplan plus system as a function of field size and depth in a water phantom for 15°, 30°, 45°, and 60° wedge filters. The attenuation coefficients, beam hardening coefficient, and beam softening coefficients were also determined experimentally using the 15 MV photon beam for each wedge angle. The measured normalized wedge factor was found to increase with increasing depth and field size for the 15 MV beam. The Theraplan plus calculated normalized wedge factor was found to be in good agreement with the experimental values. This study indicated that ignoring the dependence of the wedge factor on depth and field size will result in underexposure of the tumour.     

Determination of Boron Contents in Water Samples Collected from the Neelum valley,Azad Kashmir,Pakistan

Intake of boron from food and drinking water may pose a risk to the public health above a certain concentration level. Therefore, knowledge of boron concentration in drinking water and food items is essential. In this context, samples of drinking water were collected from natural springs of the Neelum valley, Azad Kashmir, hit by devastating earthquake in 2005. In these samples, boron concentration was determined using neutron-induced radiography technique. To do so, unknown water samples, along with standard of known boron dried on CR-39 detectors, were irradiated with thermal neutrons. After exposure, CR-39 detectors were etched in 6 M NaOH at 70°C. The tracks produced due to the alpha particles and ⁷Li ions as a result of ¹⁰B(n,α)⁷Li reaction were counted under an optical microscope. The tracks produced in theses samples were then related to the boron contents. The measured boron concentration in water samples was found to vary from 0.105 ± 0.005 to 0.247 ± 0.013 mg/l with an average value of 0.17 ± 0.04 mg/l, which are within the acceptable limits.     

Curcumin sensitizes acute promyelocytic leukemia cells to unfolded protein response-induced apoptosis by blocking the loss of misfolded N-CoR protein

Acute promyelocytic leukemia (APL) is characterized by accumulation of apoptosis-resistant immature promyelocytic cells in the bone marrow and peripheral blood. We have shown that endoplasmic reticulum (ER)-associated degradation (ERAD) and protease-mediated degradation of misfolded nuclear receptor corepressor (N-CoR) confer resistance to unfolded protein response (UPR)-induced apoptosis in APL. These findings suggest that therapeutic inhibition of N-CoR misfolding or degradation may promote growth arrest in APL cells by sensitizing them to UPR-induced apoptosis. On the basis of this hypothesis, we tested the effects of several known protein conformation-modifying agents on the growth and survival of APL cells and identified curcumin, a natural component of turmeric, as a potent growth inhibitor of APL cells. Curcumin selectively inhibited the growth and promoted apoptosis in both primary and secondary leukemic cells derived from APL. The curcumin-induced apoptosis of APL cells was triggered by an amplification of ER stress, possibly from the accumulation of misfolded N-CoR protein in the ER. Curcumin promoted this net accumulation of aberrantly phosphorylated misfolded N-CoR protein by blocking its ERAD and protease-mediated degradation, which then led to the activation of UPR-induced apoptosis in APL cells. The activation of UPR by curcumin was manifested by phosphorylation of protein kinase RNA-like endoplasmic reticulum kinase (PERK) and eukaryotic translation initiation factor 2 alpha (eIF2α), and upregulation of C/EBP homologous protein (CHOP) and GADD34, the principal mediators of proapoptotic UPR. These findings identify the therapeutic potential of curcumin in APL and further establish the rationale of misfolded N-CoR protein as an attractive molecular target in APL.     

Phosphorus and phosphate solubilizing bacteria: Keys for sustainable agriculture

Abstract

Phosphorus (P) is one of the most important minerals required for plant growth occupying a strong position among soil macro nutrients. Soil P deficiency is often fulfilled by phosphate fertilizers. P deficiency in soils is due to less total P contents in the soil and fixation of added P from chemical fertilizers as well as other organic sources like manures. The response of plant under P stress or even when it is present in adequate amount is very mild. The basic constraint in the availability of P is its solubilization as it gets fixed both in acidic and alkaline soil. Soil fixed P can only be solubilized by phosphate solubilizing microorganisms (PSMs).These bacteria released different types of organic acids in the soil which make P soluble and available to plants. The potential of these PSMs to solubilize P varies and mainly depends upon mechanism adopted for solubilization, their molecular genetics as well as their ability to release P in soil. The PSMs, having all the characteristics of phytohormone production, nitrogen fixation, as well as, heavy metal decontamination and creating salt stress tolerance in plants, are quite rare for sustainable agriculture. Application of this environment friendly approach for increasing crop productivity as well as its impact on soil and plant health is discussed in this review which will not only open new avenues of research but also provide fruitful information about phosphate solubilizing microbes for sustainable agriculture development.