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The gene lac-1, encoding the enzyme laccase, is one of several genes of the chestnut blight fungus, Cryphonectria parasitica, that are suppressed by virulence-attenuating mycoviruses of the hypovirus group. Two antagonistic regulatory pathways have been shown to govern the activity of the lac-1 promoter: a positive pathway that stimulates transcription and a negative pathway that represses transcription. We now report that these two regulatory pathways respond independently to specific changes in the nutritional environment. These newly defined conditions were used to confirm that a hypovirus suppresses the activity of the positive regulatory pathway, and to implicate calmodulin and calcineurin as components of the signal transduction cascades regulating lac-1 transcription. Significantly, lac-1 transcript accumulation was shown to be affected by amino acid availability. Further analysis revealed that transcriptional repression mediated by the negative regulatory pathway is relieved under conditions of amino acid deprivation. Thus, by blocking the positive pathway, hypovirus infection prevents increased lac-1 transcript accumulation in response to amino acid deficiency. These observations are consistent with the hypothesis that hypoviruses alter the transcriptional response of the host fungus to changes in nutrient availability.

A viral double-stranded RNA associated with virulence attenuation (hypovirulence) in the chestnut blight fungus (Cryphonectria parasitica) has been shown by DNA-mediated transformation to be responsible for transmissible hypovirulence. In addition to reduced virulence, the fungal strain harboring this virus exhibits a diverse array of characteristics, termed hypovirulence-associated traits, which distinguish it from an isogenic virus-free strain. We have investigated one of these traits, suppressed lac-1 (laccase) transcript accumulation. Two different and opposing regulatory pathways appear to govern lac-1 transcript levels in the virus-free strain: a stimulatory pathway was found to be dependent on the inositol trisphosphate (IP3) and calcium second messenger systems. A second pathway limiting transcript accumulation was shown to require ongoing protein synthesis. Additionally, changes in the level of lac-1 transcript accumulation were found to be related to modulation of promoter activity and this activity was shown to be suppressed in the virus-containing strain. We conclude that this hypovirulence-associated virus interferes with transduction of an IP3-calcium-dependent signal that is required for stimulation of lac-1 transcription. The perturbation of such signal transduction pathways by hypovirulence-associated viruses may account for the manifold symptoms associated with transmissible hypovirulence.