A global investigation of gene deletion strains that affect premature stop codon bypass in yeast, Saccharomyces cerevisiae.

Protein biosynthesis is an orderly process that requires a balance between rate and accuracy. To produce a functional product, the fidelity of this process has to be maintained from start to finish. In order to systematically identify genes that affect stop codon bypass, three expression plasmids, pUKC817, pUKC818 and pUKC819, were integrated into the yeast non-essential loss-of-function gene array (5000 strains). These plasmids contain three different premature stop codons (UAA, UGA and UAG, respectively) within the LacZ expression cassette. A fourth plasmid, pUKC815 that carries the native LacZ gene was used as a control. Transformed strains were subjected to large-scale ß-galactosidase lift assay analysis to evaluate production of ß-galactosidase for each gene deletion strain. In this way 84 potential candidate genes that affect stop codon bypass were identified. Three candidate genes, OLA1, BSC2, and YNL040W, were further investigated, and were found to be important for cytoplasmic protein biosynthesis.

Synthesis, characterization, crystal structure, and biological studies of two new Cd (II) complexes with 4'-(4-chlorophenyl)-2,2' :6',2"-terpyridine (Clphtpy).

Two new Cd(II) complexes, with the ligand 4'-(4-chlorophenyl)-2,2':6',2"-terpyridine (Clphtpy) formulated as: [Cd(Clphtpy)(NO3)2H2O] (1), and [Cd(Clphtpy)(N3)2]2 (2), have been synthesized and characterized by CHN elemental analysis as well as FT-IR, 1H NMR, absorption and emission spectroscopy, thermal analysis and analyzed structurally by X-ray single-crystal diffraction. The single crystal X-ray analysis showed that the coordination number in complex 1 and 2 were seven and six with N3O4 and N6 coordination sphere, respectively. The antibacterial activities of the synthesized complexes were tested against four gram-positive and four gram-negative bacteria. A biological study of the complexes indicated that the complex 1 exhibited very good activity against most of the tested bacteria and its activity was better than gentamicin as a standard antibiotic.

Large-scale investigation of oxygen response mutants in Saccharomyces cerevisiae.

A genome-wide screen of a yeast non-essential gene-deletion library was used to identify sick phenotypes due to oxygen deprivation. The screen provided a manageable list of 384 potentially novel as well as known oxygen responding (anoxia-survival) genes. The gene-deletion mutants were further assayed for sensitivity to ferrozine and cobalt to obtain a subset of 34 oxygen-responsive candidate genes including the known hypoxic gene activator, MGA2. With each mutant in this subset a plasmid based ß-galactosidase assay was performed using the anoxic-inducible promoter from OLE1 gene, and 17 gene deletions were identified that inhibit induction under anaerobic conditions. Genetic interaction analysis for one of these mutants, the RNase-encoding POP2 gene, revealed synthetic sick interactions with a number of genes involved in oxygen sensing and response. Knockdown experiments for CNOT8, human homolog of POP2, reduced cell survival under low oxygen condition suggesting a similar function in human cells.

Synthesis and characterization of a series of transition metal complexes with a new symmetrical polyoxaaza macroacyclic Schiff base ligand: X-ray crystal structure of cobalt(II) and nickel(II) complexes and their antibacterial properties.

A new symmetrical [N4O2] hexadentate Schiff base ligand, (E)-N-(pyridin-2-ylmethylene)-2-(3-(2-((E)-pyridin-2-lmethyleneamino)phenoxy)naphthalen-2-yloxy)benzenamine, abbreviated to L, and its complexes of Ni(II), Cu(II), Zn(II), Co(II), Cd(II) and Mn(II) have been synthesized in the presence of metal ions. The complexes were structurally characterized by elemental analyses, IR, UV-Vis, NMR and molar conductivity. The crystal structures of two complexes, [NiL(ONO2)2]·2H2O and [CoLCl2]CH3OH·0.5H2O, have been determined by a single crystal X-ray diffraction study. In these complexes, the ligand is coordinated in a neutral form via pyridine and azomethine nitrogen atoms. The metal ions complete their six coordination with two coordinated nitrate or chloride ions, forming a distorted octahedral geometry. The synthesized compounds have antibacterial activity against the three Gram-positive bacteria: Enterococcus faecalis, Bacillus cereus and Staphylococcus epid and also against the three Gram-negative bacteria: Citrobacter freundii, Enterobacter aerogenes and Salmonella typhi. The activity data show that the complexes are more potent antibacterials than the parent Schiff base.

Synthesis, spectroscopic characterization and antibacterial activity of new cobalt(II) complexes of unsymmetrical tetradentate (OSN2) Schiff base ligands.

Cobalt ion complexes with the Schiff bases, (4-X-2-{[2-(2-pyridine-2-yl-ethylsulfanyl)ethylimino]methyl}phenol (X=methoxy (OMe), phenylazo (N(2)Ph), bromo (Br), nitro (NO(2))),were synthesized and investigated by several techniques using elemental analysis (C, H, N), FTIR, electronic spectra and molar conductivity. The thermal stability of free ligands and related cobalt complexes were studied by using differential scanning calorimetry (DSC) and thermogravimetric analyses (TGA). Cyclic voltammetry indicates that the investigated cobalt complexes, under the experimental conditions, have irreversible redox behavior. The synthesized compounds have antibacterial activity against the four Gram-positive bacteria: Streptococcus pyogenes, Streptococcus agalactiae, Staphylococcus aureus and Bacillus anthracis and also against the two Gram-negative bacteria: Klebsiella pneumoniae and Pseudomonas aeruginosa. The activity data show that the parent Schiff bases are more potent antibacterials than the cobalt complexes.