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1.
J Bacteriol ; 183(15): 4509-16, 2001 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-11443085

RESUMO

Oxygen is a major determinant of both survival and mortality of aerobic organisms. For the facultative anaerobe Lactococcus lactis, oxygen has negative effects on both growth and survival. We show here that oxygen can be beneficial to L. lactis if heme is present during aerated growth. The growth period is extended and long-term survival is markedly improved compared to results obtained under the usual fermentation conditions. We considered that improved growth and survival could be due to the capacity of L. lactis to undergo respiration. To test this idea, we confirmed that the metabolic behavior of lactococci in the presence of oxygen and hemin is consistent with respiration and is most pronounced late in growth. We then used a genetic approach to show the following. (i) The cydA gene, encoding cytochrome d oxidase, is required for respiration and plays a direct role in oxygen utilization. cydA expression is induced late in growth under respiration conditions. (ii) The hemZ gene, encoding ferrochelatase, which converts protoporphyrin IX to heme, is needed for respiration if the precursor, rather than the final heme product, is present in the medium. Surprisingly, survival improved by respiration is observed in a superoxide dismutase-deficient strain, a result which emphasizes the physiological differences between fermenting and respiring lactococci. These studies confirm respiratory metabolism in L. lactis and suggest that this organism may be better adapted to respiration than to traditional fermentative metabolism.


Assuntos
Complexo de Proteínas da Cadeia de Transporte de Elétrons , Proteínas de Escherichia coli , Heme/metabolismo , Lactococcus lactis/crescimento & desenvolvimento , Anaerobiose , Proteínas de Bactérias/metabolismo , Meios de Cultura , Grupo dos Citocromos b , Citocromos/metabolismo , Fermentação , Ferroquelatase/metabolismo , Hidrogênio/metabolismo , Lactococcus lactis/metabolismo , Lactococcus lactis/fisiologia , Oxirredutases/metabolismo , Oxigênio/metabolismo , Fatores de Tempo
2.
J Biol Chem ; 276(11): 8469-74, 2001 Mar 16.
Artigo em Inglês | MEDLINE | ID: mdl-11078740

RESUMO

Cadmium is very toxic at low concentrations, but the basis for its toxicity is not clearly understood. We analyzed the proteomic response of yeast cells to acute cadmium stress and identified 54 induced and 43 repressed proteins. A striking result is the strong induction of 9 enzymes of the sulfur amino acid biosynthetic pathway. Accordingly, we observed that glutathione synthesis is strongly increased in response to cadmium treatment. Several proteins with antioxidant properties were also induced. The induction of nine proteins is dependent upon the transactivator Yap1p, consistent with the cadmium hypersensitive phenotype of the YAP1-disrupted strain. Most of these proteins are also overexpressed in a strain overexpressing Yap1p, a result that correlates with the cadmium hyper-resistant phenotype of this strain. Two of these Yap1p-dependent proteins, thioredoxin and thioredoxin reductase, play an important role in cadmium tolerance because strains lacking the corresponding genes are hypersensitive to this metal. Altogether, our data indicate that the two cellular thiol redox systems, glutathione and thioredoxin, are essential for cellular defense against cadmium.


Assuntos
Cádmio/toxicidade , Proteoma , Proteínas de Saccharomyces cerevisiae , Saccharomyces cerevisiae/efeitos dos fármacos , Cisteína/biossíntese , Proteínas de Ligação a DNA/fisiologia , Glutationa/biossíntese , Glutationa/genética , Estresse Oxidativo , Saccharomyces cerevisiae/genética , Tiorredoxina Redutase 1 , Tiorredoxina Dissulfeto Redutase/fisiologia , Fatores de Transcrição/fisiologia
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