• Korean Journal of Microbiology
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• v.42 no.3
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• pp.195-198
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• 2006

Lethal toxin is a critical virulence factor of anthrax. It is composed two protein: protective antigen (PA) and lethal factor (LF). PA binds to specific cell surface receptors and, forms a membrane channel that mediates entry of LF into the cell. LF is a zinc-dependent metalloprotease, which cleaves MKKs [MAPK (mitogen-activated protein kinase) kinases] at peptide bonds very close to their N-termini. In this study, we suggest application of cell-based assays in the early phase of drug discovery, with a particular focus on the use of yeast cells. We constructed MEK1 expression system in yeast to determine LF activity and approached cell-based assay system to screen inhibitors, in which the results covering the construction of LF-substrate in yeast expression vector, expression, and LF-mediated proteolysis of substrate were described. These results could provided the basic steps in design of cell-based assay system with the high efficiency, rapidly and easy way to screening of inhibitors.

• Molecules and Cells
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• v.26 no.1
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• pp.93-99
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• 2008

Transcriptional silencing is regulated by promoter methylation and histone modifications such as methylation and acetylation. We constructed a Schizosaccaromyces pombe reporter strain, KCT120a, to identify modifiers of transcriptional silencing, by inserting the $ura4^+$ gene into a heterochromatic telomere region. Two compounds inhibited the activity of histone deacetylases, induced acetylation of histone H3 and caused apoptotic cell death in HeLa cells. Expression of gelsolin and $p21^{waf1/cip1}$ also increased, as it does in response to HDAC inhibitors such as TSA. Therefore, these compounds appear to be potent inhibitors of HDACs, and hence potential anti-cancer drugs. Our observations suggest that a yeast cell-based assay system for transcriptional silencing may be useful for identifying histone deacetylase inhibitors and other agents affecting chromatin remodeling.

• Proceedings of the Microbiological Society of Korea Conference
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• 2002.10a
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• pp.162-172
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• 2002

Schizosaccharomyces pombe is suited for the study of cytokinesis as it divides by forming a septum in the middle of the cell at the end of mitosis. To enhance our understanding of the cytokinesis, we have carried out a genetic screen for temperature-sensitive S. pombe mutants that show defects in septum formation and cell division. Here we present the isolation and characterization of a new temperature-sensitive mutant, sun1(septum uncontrolled), which undergoes uncontrolled septation during cell division cycle at restrictive temperature $(37^{\circ}C)$. In sun1 mutant, actin ring and septum are positioned at random locations and angles, and nuclear division cycle continues. These observations suggest that the sun] gene product is required for the proper placement of the actin ring as well as precise septation. The sun] mutant is monogenic recessive mutation unlinked to previously known various cdc genes of S. pombe. In a screen for $sunl^+$ gene to complement the sun] mutant, we have cloned a gene, $susl^+$(suppressor of sun1 mutant), that encodes a protein of 689 amino acids. The predicted amino acid sequence of $susl^+$ gene is similar to the human hMadlp and Saccharomyces cerevisiae Mad1p, a component of the spindle checkpoint in eukaryotic cells. The null mutant of $susl^+$ gene grows normally at various temperatures and has the increased sensitivity to anti-microtubule drug, while $susl^+$ mutant shows no sensitivity to microtubule destabilizing drugs. The putative S. pombe Sus1p directly interacts with S. pombe Mad2p in yeast two-hybrid assays. These data suggest that the newly isolated susr gene encodes S. pombe Mad1p and suppresses sun] mutant defective in controlled septation in a cell division cycle.

• Advances in nano research
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• v.13 no.4
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• pp.325-339
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• 2022

Endophytes ascertain a symbiotic relationship with plants as promoters of growth, defense mechanism etc. This study is a first report to screen the endophytic population in Waltheria indica, a tropical medicinal plant. 5 bacterial and 3 fungal strains in leaves, 3 bacterial and 1 yeast species in stems were differentiated morphologically and identified by biochemical and molecular methods. The phylogenetic tree of the isolated endophytes was constructed using MEGA X. Silver nanoparticles were biosynthesized from a rare endophytic bacterium Cupriavidus metallidurans isolated from the leaf of W. indica. The formation of silver nanoparticles was confirmed by UV-Visible spectrophotometer that evidenced a strong absorption band at 408.5 nm of UV-Visible range with crystalline nature and average particle size of 16.4 nm by Particle size analyzer. The Fourier Transform Infra-Red spectrum displayed the presence of various functional groups that stabilized the nanoparticles. X-ray diffraction peaks were conferred to face centered cubic structure. Transmission Electron Microscope and Scanning Electron Microscope revealed the spherical-shaped, polycrystalline nature with the presence of elemental silver analyzed by Energy Dispersive of X-Ray spectrum. Selected area electron diffraction also confirmed the orientation of AgNPs at 111, 200, 220, 311 planes similar to X-ray diffraction analysis. The synthesized nanoparticles are evaluated for antimicrobial activity against 7 bacterial and 3 fungal pathogens. A good zone of inhibition was observed against pathogenic bacteria than fungal pathogens. Thus the study could hold a key aspect in drug discovery research and other pharmacological conducts of human clinical conditions.