TABLE 3.

V-ATPase activity, cyclosporin A sensitivity, and quinacrine staining of candidatevma mutants

Strain	% wild type ATPase activity	CsA sensitive?	Quinacrine
Wild type	100	−pH 5/−pH 6	++
vma3Δ	5a	+pH 5/+pH 6	−
kex2Δ	103b	+pH 5/+pH 6	++
rib4Δ	ND	+pH 5/+pH 6	++
pho85Δ	85 (1)	−pH 5/+pH 6	++
snf5Δ	80 (1)	+pH 5/+pH 6	++
snf6Δ	60 (1)	−pH 5/−pH 6	++
rcs1Δ	ND	−pH 5/+pH 6	++
anp1Δ	90 (1)	−pH 5/−pH 6	++
ctr1Δ	ND	−pH 5/−pH 6	++
cwh36Δ	5 ± 2 (2)	+pH 5/+pH 6	−
vps34Δ	17 ± 7 (2)	−pH 5/+pH 6	+
vps15Δ	15 ± 5 (2)	−pH 5/+pH 6	+
vps45Δ	32 ± 10 (2)	−pH 5/−pH 6	+
vps16Δ	1 ± 1 (2)	+pH 5/+pH 6	++?d
shp1Δ	28 ± 9 (2)	+pH 5/+pH 6	+
clc1Δ	23 ± 12 (2)	+pH 5/+pH 6	++?d
rnr1Δ	35 ± (2)	−pH 5/+pH 6	+
rmd7Δ	28 ± 3 (2)	−pH 5/−pH 6	+?d
cys4Δ	47c	+pH 5/ +pH 6	+

Comparison of V-ATPase activity is shown in isolated vacuolar membranes with quinacrine staining and cyclosporin A sensitivity. Vacuolar membranes were isolated from the BY4741 wild-type strain and from the indicated mutant strains, and ATPase activity sensitive to 100 nm concanamycin A was determined as described in materials and methods. The wild-type vacuolar vesicles had an average concanamycin-sensitive specific activity of 2.0 ± μmol Pi/min/mg protein when assayed at 37° (average of n = 7 independent vacuole isolations ±SEM). Activities from the mutant vacuole preparations are expressed as a percentage of the wild-type activity ± the range of duplicate assays where indicated. CsA sensitivity and quinacrine staining are derived from Figures 3 and 2, respectively.

^aFrom Umemoto et al. (1990).

^bFrom Oluwatosin and Kane (1998).

^cFrom Oluwatosin and Kane (1997).

^dThese strains show either such perturbed vacuolar morphology (vps16Δ and clc1Δ) or a variability in staining among different cells (rmd7Δ) that assessment of quinacrine staining is difficult.