TABLE 2.

Disassembly and reassembly of V-ATPase in mutant strains^a

Strain	% of 100-kDa subunit assembled into V1V0 complexes
	In YEPD	After YEP treatment	After glucose readdition
SF838-1D	77.5 ± 11.5	18.6 ± 3.6	74.5 ± 14.4
W303-1A snf1Δ	52.9	12.3	62.6
W303-1A rts1Δ	42.3	13.4	62.6
W303-1A	49.2 ± 0.8	13.6 ± 6.4	40.7 ± 0.7
DL100-1783 pkc1Δ	77.4 ± 4.9	19.2 ± 0.2	70.5 ± 8.9
DL100-1783	71.8 ± 9.1	20.2 ± 1.9	60.0 ± 5.0
SF838-5A [rho0]	63.6	19.5	60.7
W303-1A pde2Δ
No added cAMP	56.3	21.2	55.5
+5 mM cAMP with YEPD	53.4	13.7
+5 mM cAMP with YEP		15.9
+5 mM cAMP after 5 min with YEP		21.5

^aMutant strains and their isogenic wild-type strains were labeled with Tran³⁵S-label and chased in YEPD, YEP, or YEP followed by addition of 2% glucose. V₁V₀ and free V₀ complexes were immunoprecipitated by antibodies 8B1 and 10D7, respectively. The percentage of 100-kDa subunit assembled into V₁V₀ complexes was calculated by quantitating the amount immunoprecipitated by each antibody on a PhosphorImager and dividing the amount immunoprecipitated by antibody 8B1 by the total immunoprecipitated by both antibodies. SF838-1D is the wild-type strain used throughout the rest of this work, and it is included for comparison. W303-1A is the isogenic wild-type strain for the snf1Δ, rts1Δ, and pde2Δ mutants, and DL100-1783 is the isogenic wild-type strain for the pkc1Δ mutant. The genetic background of the [rho⁰] strain (SF838-5A) is very similar to that of SF838-1D and gave qualitatively similar results in disassembly and reassembly experiments. Results are expressed as mean ± standard deviation for SF838-1D (n = 5) and mean ± range of two experiments for W303-1A, DL100-1783, and DL100-1783 pkc1Δ. Quantitation for the other strains was from a single experiment.