FIG. 1.

Disruption of the adenylyl cyclase gene CAC1 abolishes cAMP production. (A) CAC1 wild-type (WT) (H99), cac1 mutant (RPC3), cac1 CAC1-reconstituted (RPC7), and gpa1 CAC1 (AAC17) strains were incubated in YPD medium for 18 h. The cells were divided for incubation for 4 h in one of three different media: synthetic complete medium with 2% glucose (nitrogen +, glucose +), synthetic complete medium with 0% glucose (nitrogen +, glucose −), or SLAD (nitrogen −, glucose +). Total RNA from each sample was assessed by Northern analysis with the CAC1 gene as a probe. The rRNA bands of the ethidium bromide-stained gel (rRNA) are shown to demonstrate RNA loading. (B) CAC1 wild-type (H99) (squares), cac1 mutant (LCC22) (diamonds), cac1 CAC1-reconstituted (LCC22-1) (triangles), gpa1 mutant (AAC1) (circles), gpa1 GPA1-reconstituted (AAC3) (crossed squares), and gpa1 CAC1 (AAC17) (crosses) strains were starved for glucose for 2 h. At the indicated time after a glucose pulse, aliquots of the cell suspensions were frozen, and intracellular cAMP concentrations were determined. Data points represent the mean and standard deviation for duplicate samples in two identical experiments (four samples for each data point).