Many extracellular signals trigger changes in gene expression by altering the steady-state level of target transcripts. This modulation of transcript levels is typically ascribed to changes in transcription of target genes; however, there are numerous examples of changes in mRNA processing and stability that contribute to the overall change in transcript levels following signalling pathway activation. The alpha-factor-stimulated mating pathway in Saccharomyces cerevisiae is a receptor-operated MAP kinase cascade that results in increased levels of a large number of target mRNA transcripts when stimulated acutely. A previous study identified many of the transcripts modulated in response to alpha-factor and argued, based on genetic studies, that the response occurred solely at the level of gene transcription (Roberts et al., 2000). We directly examined whether enhanced mRNA stability contributes to the increase in the steady-state level of alpha-factor target transcripts by exploiting a temperature-sensitive RNA Polymerase II mutant, a Ste12 transcription factor import mutant, and tet-regulated synthetic mating factor minigene reporters. Examination of a panel of alpha-factor-responsive transcripts reveals no change in mRNA stability in response to alpha-factor stimulation, providing direct evidence that this signal transduction pathway in S. cerevisiae does not function through modulating transcript stability.