Journal of Advances in Molecular Biology

19

Authors: John M. Schmitt, Jessica Magill, Amanda Ankeny, Renee Geck, Jessica Milligan, Hannah McFarland, Erica Rice

Calcium/calmodulin-dependent protein kinase (CaM Kinase) proteins are targets of hormones and growth factors and regulate cancer cell growth, apoptosis and migration. The hormone estrogen (E2) utilizes CaM Kinases to activate the Extracellular-signal regulated kinase (ERK) leading to MCF-7 breast cancer cell growth. The hormone Vitamin D (Vit D) may inhibit breast cancer cell growth however the cellular mechanisms of Vit D action remain to be elucidated. Within the present study we provide data that E2 stimulation of MCF-7 cells activates CaM Kinase Kinase (CaM KK), CaM KI, and ERK and the transcription factors Elk-1 and SRF. E2 treatment of MCF-7 cells potently stimulated Elk-1/SRF directed transcription of SRE-luciferase reporters. E2 activation of ERK, Elk-1, SRF and SRE-dependent luciferase activities were blocked by treating cells with the CaM KK inhibitor, STO-609, and the ERK inhibitor, U0126. Moreover, siRNAs directed against CaM KK and ERK blocked transcription factor phosphorylation and luciferase activity. Treatment of cells with Vit D, the hormone Epinephrine (Epi), or Forskolin, prevented E2 activation of CaM KK and ERK. Interestingly, Vit D promoted a PKA-dependent phosphorylation and inhibition of CaM KK as well as its association with 14-3-3. Epi and Vit D treatment of cells blocked the ability of E2 to active Elk-1 and SRE-luciferase activity. This data suggests an important role for CaM KK and ERK in regulating transcription downstream of E2 in MCF-7 cells. Our results also suggest that Vit D treatment of MCF-7 cells utilizes a unique PKA-dependent mechanism to block E2 activation of CaM KK, ERK and transcription.

7

Authors: Anicet Batcho, Bilal Sarwa, Bushra Rachid

Identifying active domain in a protein sequence is one of the most important functions in molecular biology. This is usually studied with three-dimensional (3D) structure of the active sites protein using Bioinformatics tools. In the absence of an experimentally determined structure, homology modeling can sometimes supply a useful 3D model for protein that is linked to at least one known protein structure. In this work, we have evaluated the sequence information of Heat Shock Protein (HSP) gene retrieved from Agave americana. Homology modeling, functional annotation of A. americana heat shock protein (HSP) sequence. Based on homology modeling, 3D structure of the gene was constructed. Validation tests were used to calculate and determine the reliability of the structure. In this study, details structure of HSP led to develop a better understanding of the HSP’s role in different abiotic stress response pathways.

8

Authors: Abu Bakr El-Bediwi, El-Sayed Kamel Areida, Eman Eid

The present study aimed to investigate morphology structure, blood structure and the rheological properties, (viscosity, torque, shear stress and shear rate), of mice blood serum at wide range of shear rates after whole body exposed to different ultraviolet doses. Eighty healthy male mice were used in this study. A significant increase in the mice blood serum viscosity occurs after irradiated by the ultraviolet C radiation (UVC). The relationship between viscosity and shear rate exhibited a non-Newtonian behavior for all mice group. The relationship between shear stress and % torque and shear rate exhibited a linear behavior for the control and UVC irradiated mice group. The mice blood serum showed a non-significant change in the UVC irradiated mice % torque and shear stress with shear rates (40-500 s-1) compared with the control. There was a noticeable change in the general behavior movement with a significant effect in blood mice structure after exposure to UVC radiation.

14

Authors: Saranya. A., Sivakumari. K., Ashok. K., Rajesh. S

The present study aims at the phytochemical profiling of lyophilized pure fruit juice powder of Citrus limon and demonstrates its efficacy as an antitumor agent against human breast cancer cell line (MCF-7). The cytotoxicity was evaluated using the changes in cell morphology, cell viability, and DNA fragmentation, the percentage of cell viability was determined by MTT assay. Our results showed that lyophilized pure fruit juice powder of Citrus limon inhibited the proliferation of human breast cancer cell line MCF-7 with an IC50 98.16 μg/ml at 48 h incubation, it was shown to promote apoptosis as seen as DNA fragmentation using ladder assay. These results suggest that lyophilized pure fruit juice powder of Citrus limon has antiproliferative effect against MCF-7 cell by suppressing its growth.

8

Authors: Manju T, Anju T R

Hepatic tissue damage mediated through altered antioxidant defence is one of the characteristic features of diabetes. Phytotherapy using Aegle marmelos leaf extract has been reported to control diabetes complications on brain functions. The present study was carried out to evaluate whether the anti-hypergycemic effects of Aegle marmelos leaf extract can regulate diabetes related hepatic oxidative stress. To understand this, antioxidant and peroxide levels and expression of stress related Hypoxia inducible factor 1 (HIF 1) mRNA were analysed in liver tissue of experimental rat models. The decreased total antioxidant level with elevated peroxide level along with upregulation of HIF 1 gene expression indicates poor oxidative defenses in the liver of diabetic rats. These altered parameters were reversed to near control level by Aegle marmelos treatment which indicates that Aegle marmelos can not only regulate blood glucose levels but also control diabetes related hepatic complications.

19

Authors: Moncef Boulila

Recombination plays a key role in virus evolution. Except a limited number of works carried out so far on recombination in Prunus necrotic ringspot virus (PNRSV) genome, knowledge is still poor. Numerous phylogenetic studies particularly those based on capsid (CP) and movement protein (MP) of PNRSV showed that all accessions described so far split into three major phylogroups, i.e., PV96, PV32, and PE5. With the recent characterization of Chilean and Indian isolates which were phylogenetically closely related to the oldest member, i.e., SW6 (AF013287) from USA, a new phylogroup baptized SW6, emerged since it was outside of the evolutionary behavior of the classical PV96, PV32, and PE5 clusters. In order to understand how it evolved, six computerized methods were used to detect potential recombination in the CP sequences of nine components forming SW6 phylogroup. It was clearly demonstrated that various members underwent recombination. Additionally, networked relationships among them proved that numerous incompatibilities occurred in these sequences illustrated by the presence of boxes in the network implying therefore the possibility of recombination. According to inferred network, several sequences contained conflicting signals as illustrated by various splits and edges with special reference to Chilean (EF565255, EF565256), polish (AF332614) and Indian (AM494934) isolates. It is worth noting that the phylogenetic network provided a different clustering compared to the bifurcating tree. In fact, three distinct groups were delineated with SplitsTree4 software instead of a globally homogenous ensemble generated by MEGA7 algorithm. Furthermore, this study pointed out that members of SW6 phylogroup were the only recombinants among 205 tested accessions confirming that SW6 should be considered as the fourth phylogenetic group of PNRSV. On the other hand, CP was predominantly under purifying selection. However, positive selection was evidenced particularly in the C-terminal region of CP comprising part of dimerization region (DR). Such adaptive selection was exerted on a CP segment of AM494934 accession exchanged by recombination.

22

Authors: D. Gupta, K. Bhardwaj, R. Gothalwal, S. Bhargava

In this paper, we examined the effect of salt stress (NaCl) and osmotic stress (sucrose) on proteomic level in the diazotrophic cyanobacterium Nostoc muscorum. The aim of this study is to compare proteins appeared in control vs. salt treated, control vs. sucrose treated and salt treated vs. sucrose treated cultures. In the salt treated cultures about 37 proteins were expressed differentially out of these only 5 proteins have shown fold regulation of 1.5 or more. About 141 proteins were found to express independently in control and about 554 proteins were express independently in salt treated culture. When we compared proteins in control and sucrose treated cells, it was reported that about 37 protein spots were express differentially, out of these only 7 proteins have fold regulation 1.5 or more. The independently expressed proteins appeared on gel are 141 and 186 respectively. Similarly, when we compared proteins appeared in salt and sucrose treated cells, it was reported that about 54 proteins were express differentially, out of these 10 proteins have fold regulation 1.5 or more. About 537 protein spots were independently present in salt treated cells and about 186 proteins were independently present in sucrose treated cells. In addition, the differentially expressed proteins and their identification with their functional group have also been discussed.