The role of oxygen and reduced oxygen species in nitric oxide-mediated cytotoxicity: studies in the yeast Saccharomyces cerevisiae model system.

The cytotoxicity of nitric oxide (NO) is well established, yet the mechanism(s) of its cytotoxicity is (are) still undefined and a matter of significant interest and speculation. Many of the previously proposed mechanisms for NO-mediated cytotoxicity involve interactions between NO and molecular oxygen (O(2)) and/or O(2)-derived species such as O(-)(2) and H(2)O(2). The yeast Saccharomyces cerevisiae represents a useful model system for evaluating the role of O(2) and O(2)-derived species in NO-mediated cytotoxicity. This study examines the contribution of O(2) and O(2)-derived species to NO-mediated cytotoxicity in the yeast S. cerevisiae. NO-mediated cytotoxicity was determined to be O(2)-dependent. However, this O(2) dependence was only minimally due to the generation of O(2)-derived species such as O(-)(2) and/or H(2)O(2).

Effects of nitric oxide on the copper-responsive transcription factor Ace1 in Saccharomyces cerevisiae: cytotoxic and cytoprotective actions of nitric oxide.

Previous studies indicate that nitric oxide (NO) can serve as a regulator/disrupter of metal-metabolizing systems in cells and, indeed, this function may represent an important physiological and/or pathophysiological role for NO. In order to address possible mechanisms of this aspect of NO biology, the effect of NO on copper metabolism and toxicity in the yeast Saccharomyces cerevisiae was examined. Exposure of S. cerevisiae to NO resulted in an alteration of the activity of the copper-responsive transcription factor Acel. Low concentrations of the NO donor DEA/NO were found to slightly enhance copper-mediated activation of Acel. Since Acel regulates the expression of genes responsible for the protection of S. cerevisiae from metal toxicity, the effect of NO on the toxicity of copper toward S. cerevisiae was also examined. Interestingly, low concentrations of NO were also found to protect S. cerevisiae against the toxicity of copper. The effect of NO at high concentrations was, however, opposite. High concentrations of DEA/NO inhibited copper-mediated Acel activity. Correspondingly, high concentrations of DEA/NO (1 mM) dramatically enhanced copper toxicity. An intermediate concentration of DEA/NO (0.5 mM) exhibited a dual effect, enhancing toxicity at lower copper concentrations (<0.5 mM) and protecting at higher (> or =0.5 mM) copper concentrations. Thus, it is proposed that the ability of NO to both protect against (at low concentrations) and enhance (at high concentration) copper toxicity in S. cerevisiae is, at least partially, a result of its effect on Acel. The results of this study have implications for the role of NO as a mediator of metal metabolism.

The interaction of nitric oxide (NO) with the yeast transcription factor Ace1: A model system for NO-protein thiol interactions with implications to metal metabolism.

Nitric oxide (NO) was found to inhibit the copper-dependent induction of the yeast CUP1 gene. This effect is attributable to an inhibition of the copper-responsive CUP1 transcriptional activator Ace1. A mechanism is proposed whereby the metal binding thiols of Ace1 are chemically modified via NO- and O(2)-dependent chemistry, thereby diminishing the ability of Ace1 to bind and respond to copper. Moreover, it is proposed that demetallated Ace1 is proteolytically degraded in the cell, resulting in a prolonged inhibition of copper-dependent CUP1 induction. These findings indicate that NO may serve as a disrupter of yeast copper metabolism. More importantly, considering the similarity of Ace1 to other mammalian metal-binding proteins, this work lends support to the hypothesis that NO may regulate/disrupt metal homeostasis under both normal physiological and pathophysiological circumstances.

The effect of nonionic detergents on the activity and/or stability of rat brain nitric oxide synthase.

The results of this study indicate that the addition of low concentrations of a nonionic detergent such as those represented by the Tween, Brij, or Triton classes causes an apparent activation of nitric oxide synthase. It is possible that this apparent activation is due to the ability of these detergents to stabilize the protein. The stabilizing influence of the detergents may be a result of inhibiting the dissociation of the dimeric protein into monomers or the dissociation of an essential cofactor or prosthetic group from the active enzyme. Regardless of the mechanism of action, the addition of low concentrations of nonionic detergents results in longer and increased nitric oxide synthase activity and may be an important tool for those involved in enzymological studies of nitric oxide synthase.

N,O-diacylated-N-hydroxyarylsulfonamides: nitroxyl precursors with potent smooth muscle relaxant properties.

N,O-Diacylated-N-hydroxyarylsulfonamides are capable of slowly releasing nitroxyl (HNO) by simple, non-enzymatic hydrolysis in Krebs solution at 37 degrees C. Release of nitric oxide (NO) was not seen. These compounds were also found to elicit vasorelaxation in rabbit thoracic aorta in vitro, presumably as a result of their ability to release HNO. This effect was enhanced by the addition of superoxide dismutase (SOD). Thus, these results are consistent with previous work indicating that HNO is a potent vasorelaxant.