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N-Acetyltransferase Mpr1 of Saccharomyces cerevisiae can reduce intracellular oxidation levels and protect yeast cells under oxidative stress, including H(2)O(2), heat-shock, or freeze-thaw treatment. Unlike many antioxidant enzyme genes induced in response to oxidative stress, the MPR1 gene seems to be constitutively expressed in yeast cells. Based on a recent report that ethanol toxicity is correlated with the production of reactive oxygen species (ROS), we examined here the role of Mpr1 under ethanol stress conditions. The null mutant of the MPR1 and MPR2 genes showed hypersensitivity to ethanol stress, and the expression of the MPR1 gene conferred stress tolerance. We also found that yeast cells exhibited increased ROS levels during exposure to ethanol stress, and that Mpr1 protects yeast cells from ethanol stress by reducing intracellular ROS levels. When the MPR1 gene was overexpressed in antioxidant enzyme-deficient mutants, increased resistance to H(2)O(2) or heat shock was observed in cells lacking the CTA1, CTT1, or GPX1 gene encoding catalase A, catalase T, or glutathione peroxidase, respectively. These results suggest that Mpr1 might compensate the function of enzymes that detoxify H(2)O(2). Hence, Mpr1 has promising potential for the breeding of novel ethanol-tolerant yeast strains.

N-Acetyltransferase Mpr1 of Saccharomyces cerevisiae can reduce intracellular oxidation levels and protect yeast cells under oxidative stress. We found that yeast cells exhibited increased levels of reactive oxygen species during freezing and thawing. Gene disruption and expression experiments indicated that Mpr1 protects yeast cells from freezing stress by reducing the intracellular levels of reactive oxygen species. The combination of Mpr1 and l-proline could further enhance the resistance to freezing stress. Hence, Mpr1 as well as l-proline has promising potential for the breeding of novel freeze-tolerant yeast strains.

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1. The modulation of dog atrial swelling‐induced chloride current (I(Cl,swelling)) by cAMP‐elevating agents was studied. Forskolin (10 microM) or isoprenaline (1 microM) exerted multiple effects. Although the pattern between cells was variable, there was, in general, a stimulatory action and a more slowly developing inhibitory effect. 2. In any given cell, the response to forskolin or isoprenaline was qualitatively similar suggesting that all of the responses were dependent on stimulation of adenylyl cyclase. The effects of forskolin or isoprenaline on I(Cl,swelling) were inhibited by intracellular dialysis with a P‐site inhibitor of adenylyl cyclase, 2'‐deoxyadenosine 3'‐monophosphate (300 microM). 3. Intracellular dialysis with a peptide inhibitor of protein kinase A (PKI(6‐22); 100 microM) blocked the inhibitory response to forskolin or isoprenaline and all cells responded with a monophasic stimulation of I(Cl,swelling). 4. After intracellular dialysis of cells with PKI(6‐22) (100 microM) and cAMP (100 microM), current amplitude was not further stimulated by forskolin. 5. After intracellular dialysis with PKI(6‐22) and adenosine 5'‐O‐(3‐thiotriphosphate) (ATPgammaS), forskolin stimulated I(Cl,swelling) and the effect of forskolin subsided after it was washed out. 6. In conclusion, there are dual pathways by which cAMP can modulate dog atrial cell I(Cl,swelling). Inhibition results from protein kinase A (PKA)‐dependent phosphorylation. In addition, a stimulatory pathway exists that is independent of phosphorylation by PKA or other cellular kinases. Although alternative explanations are possible, the stimulatory effect of cAMP may represent a direct modulation of I(Cl,swelling).

It has been recently reported that 5-hydroxytryptamine (5-HT) increases force of contraction in atrial tissue but not in ventricular tissue. In the present study with trabeculae obtained from non-diseased human hearts, we investigated whether this difference in the contractile responses is specific for 5-HT or is also observed for other substances: calcitonin gene-related peptide (CGRP), angiotensin II, adenosine, somatostatin and acetylcholine. CGRP (10(-9) to 10(-7) M) and angiotensin II (10(-9) to 10(-5) M) caused concentration-dependent increases in force of contraction in atrial trabeculae (up to 36 +/- 8% and 42 +/- 8% of the response to 10(-5) M noradrenaline, respectively). Similar to 5-HT, no effects were observed with CGRP and angiotensin II in ventricular trabeculae. Adenosine (10(-8) to 10(-5) M) and somatostatin (10(-8) to 10(-6) M) caused concentration-dependent negative inotropic effects on baseline atrial contractility (-54 +/- 17% and -51 +/- 25%, respectively), but no response was found on baseline ventricular contractility. Adenosine, but not somatostatin, reduced force of contraction after pre-stimulation with 10(-5) M noradrenaline in atrial tissue and, to a lesser extent, in ventricular tissue. Acetylcholine exhibited a biphasic concentration-response curve in the atrial tissue, consisting of an initial negative inotropic response (10(-9) to 10(-7) M, from 120 +/- 41 mg at baseline to 48 +/- 16 mg at 10(-7) M), followed by a positive inotropic response (10(-6) to 10(-3) M, from 48 +/- 16 mg at 10(-7) M to 77 +/- 15 mg).(ABSTRACT TRUNCATED AT 250 WORDS)

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