Arthrobacter sp. KMSZP6 isolated from a pristine uranium ore deposit at Domiasiat located in North-East India exhibited noteworthy tolerance for cesium (Cs) and strontium (Sr). The strain displayed a high minimum inhibitory concentration (MIC) of 400 mM for CsCl and for SrCl2. Flow cytometric analysis employing membrane integrity indicators like propidium iodide (PI) and thiazole orange (TO) indicated a greater sensitivity of Arthrobacter cells to cesium than to strontium. On being challenged with 75 mM of Cs, the cells sequestered 9612 mg Cs g−1 dry weight of cells in 12 h. On being challenged with 75 mM of Sr, the cells sequestered 9989 mg Sr g−1 dry weight of cells in 18 h. Heat killed cells exhibited limited Cs and Sr binding as compared to live cells highlighting the importance of cell viability for optimal binding. The association of the metals with Arthrobacter sp. KMSZP6 was further substantiated by Field Emission-Scanning Electron Microscopy (FE-SEM) coupled with Energy dispersive X-ray (EDX) spectroscopy. This organism tolerated up to 1 kGy 60Co-gamma rays without loss of survival. The present report highlights the superior tolerance and binding capacity of the KMSZP6 strain for cesium and strontium over other earlier reported strains and reveals its potential for bioremediation of nuclear waste.Electronic supplementary materialThe online version of this article (doi:10.1186/s13568-016-0247-3) contains supplementary material, which is available to authorized users.
TOR (target of rapamycin) protein kinase acts as a central controller of cell growth and development of an organism. Present study was undertaken to find the expression pattern and role of TOR during growth and development of Dictyostelium discoideum. Failures to generate either knockout and/or knockdown mutants indicate that interference with its levels led to cellular defects. Thus, the effects of TOR (DDB_G0281569) overexpression specifically, cells expressing Dd(Δ211-TOR)-Eyfp mutant was analyzed. Elevated expression of (Δ211-TOR)-Eyfp reduced both cell size and cell proliferation. DdTOR was found to be closer to fungus. mRNA level of TOR was found maximally in the freshly starved/aggregate cells that gradually declined. This was also strengthened by the expression patterns observed by in situ and the analysis of β-galactosidase reporter driven by the putative TOR promoter. The TOR protein was found to be highest at the aggregate stage. The fusion protein, (Δ211-TOR)-Eyfp was localized to the cell membrane, cytosol, and the nucleus. We suggest, DdTOR to be an essential protein and high TOR expression inhibits cell proliferation.
Dictyostelium discoideum encodes a single Rheb protein showing sequence similarity to human homologues of Rheb. The DdRheb protein shares 52 percent identity and 100 percent similarity with the human Rheb1 protein. Fluorescence of Rheb yellow fluorescent protein fusion was detected in the D. discoideum cytoplasm. Reverse transcription-polymerase chain reaction and whole-mount in situ hybridization analyses showed that rheb is expressed at all stages of development and in prestalk cells in the multicellular structures developed. When the expression of rheb as a fusion with lacZ was driven under its own promoter, the beta-galactosidase activity was seen in the prestalk cells. D. discoideum overexpressing Rheb shows an increase in the size of the cell. Treatment of the overexpressing Rheb cells with rapamycin confirms its involvement in the TOR signalling pathway.
Active DNA-dependent ATPase A Domain inhibitor (ADAADi) is the only known inhibitor of ATP-dependent chromatin remodeling proteins that targets the ATPase domain of these proteins. The molecule is synthesized by aminoglycoside phosphotransferase enzyme in the presence of aminoglycosides. ADAADi interacts with ATP-dependent chromatin remodeling proteins through motif Ia present in the conserved helicase domain, and thus, can potentially inhibit all members of this family of proteins. We show that mammalian cells are sensitive to ADAADi but with variable responses in different cell lines. ADAADi can be generated from a wide variety of aminoglycosides; however, cells showed differential response to ADAADi generated from various aminoglycosides. Using HeLa and DU145 cells as model system we have explored the effect of ADAADi on cellular functions. We show that the transcriptional network of a cell type is altered when treated with sub-lethal concentration of ADAADi. Although ADAADi has no known effects on DNA chemical and structural integrity, expression of DNA-damage response genes was altered. The transcripts encoding for the pro-apoptotic proteins were found to be upregulated while the anti-apoptotic genes were found to be downregulated. This was accompanied by increased apoptosis leading us to hypothesize that the ADAADi treatment promotes apoptotic-type of cell death by upregulating the transcription of pro-apoptotic genes. ADAADi also inhibited migration of cells as well as their colony forming ability leading us to conclude that the compound has effective anti-tumor properties.
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