Protective role of mitochondrial superoxide dismutase against high osmolarity, heat and metalloid stress in saccharomyces cerevisiae.

Superoxide dismutases, both cytosolic Cu, Zn-SOD encoded by SOD1 and mitochondrial Mn-SOD encoded by SOD2, serve Saccharomyces cerevisiae cells for defense against the superoxide radical but the phenotypes of sod1A and sod2delta mutant strains are different. Compared with the parent strain and the sod1delta mutant, the sod2delta mutant shows a much more severe growth defect at elevated salt concentrations, which is partially rescued by 2 mmol/L glutathione. The growth of all three strains is reduced at 37 degrees C, the sod2delta showing the highest sensitivity, especially when cultured in air. Addition of 1 mmol/L glutathione to the medium restores aerobic growth of the sod1delta mutant but has only a minor effect on the growth of the sod2delta strain at 37 degrees C. The sod2delta strain is also sensitive to AsIIl and AsV and its sensitivity is much more pronounced under aerobic conditions. These results suggest that, unlike the Sodlp protein, whose major role is oxidative stress defense, Sod2p also plays a role in protecting S. cerevisiae cells against other stresses--high osmolarity, heat and metalloid stress.

Effect of antioxidants on Saccharomyces cerevisiae mutants deficient in superoxide dismutases.

S. cerevisiae strain delta sodl lacking Cu,Zn-superoxide dismutase and delta sodl delta sod2 mutant lacking both Cu,Zn-SOD and Mn-superoxide dismutase displayed strongly reduced aerobic growth on glucose, glycerol and lactate; delta sod2 deletion had no effect on aerobic growth on glucose and largely precluded growth on glycerol and lactate. The oxygen-induced growth defects and their alleviation by antioxidants depended on growth conditions, in particular on oxygen supply to cells. Under strong aeration, vitamins A and E had a low effect, 100 mumol/L quercetin alleviated the growth defects of all three mutants while beta-carotene had no growth-restoring effect. The superoxide producer paraquat inhibited the aerobic growth of all three mutants in a concentration-dependent manner. Low concentrations of antioxidants had no effect on paraquat toxicity while higher concentrations supported the toxic effect of the agent.

Purification and characterization of the DNA cleavage and recognition site of I-ScaI mitochondrial group I intron encoded endonuclease produced in Escherichia coli.

The second intron in the mitochondrial cytb gene of Saccharomyces capensis, belonging to group I, encodes a 280 amino acid protein containing two LAGLIDADG motifs. Genetic and molecular studies have previously shown that this protein has a dual function in the wild-type strain. It acts as a specific homing endonuclease I- Sca I promoting intron mobility and as a maturase promoting intron splicing. Here we describe the synthesis of a universal code equivalent to the mitochondrial sequence coding for this protein and the in vitro characterization of I- Sca I endonuclease activity, using a truncated mutant form of the protein p28bi2 produced in Escherichia coli. We have also determined the cleavage pattern as well as the recognition site of p28bi2. It was found that p28bi2 generates a double-strand cleavage downstream from the intron insertion site with 4 nt long 3'-overhangs. Mutational analysis of the DNA target site shows that p28bi2 recognizes a 16-19 bp sequence from positions -11 to +8 with respect to the intron insertion site.

The detection of Salmonella in skimmed milk powder enrichments using conventional methods and immunomagnetic separation.

Skimmed milk powders were spiked with one of three Salmonella serovars and incubated in buffered peptone water for 24 h. No false-negative results were obtained by immunomagnetic separation (IMS), compared to seven for selenite cysteine, one for Müller-Kauffmann tetrathionate and two for Rappaport-Vassiliadis enrichment broths. Salmonella virchow was detected and enumerated during the pre-enrichment incubation by IMS and indirect conductance techniques. The Salm. virchow cell number did not increase after 12 h incubation and remained at 3 x 10(6) cfu ml-1. IMS was able to capture Salm. virchow cells at cell numbers ca 50 ml-1 in the presence of a 1000 greater number of non-salmonella cells.

Population dynamics in the co-culture of Shigella flexneri 1b original strain and its antigenic 3b mutant carrying a prophage.

The antigenic mutant Shigella flexneri 3b showed selective prevalence when subcultured with the original strain 1b. Mathematical analysis of such co-cultures showed that the dynamics of bacterial growth may be described by equations of the Lotka-Volterra type. The analysis of serial cultivations suggests that parameters of the equations may be realizations of random variables characterizing strains and media. The mutant carries a prophage lethal for the original strain. Distinctive features of the growth of these strains alone and in co-culture may be successfully explained by the differences in growth parameters, without phage inference. The concurrence model connected only with the existence of the phage is not sufficient. A complete description of the dynamics of the co-culture is obtained by the connection of the assumptions of two models: strains in competition and growth of strains with phage, given the random character of the parameters.