CALCINEURIN-DEPENDENT GROWTH OF AN FK506-HYPERSENSITIVE AND CSA-HYPERSENSITIVE MUTANT OF SACCHAROMYCES-CEREVISIAE

被引:54
作者
PARENT, SA [1 ]
NIELSEN, JB [1 ]
MORIN, N [1 ]
CHREBET, G [1 ]
RAMADAN, N [1 ]
DAHL, AM [1 ]
HSU, MJ [1 ]
BOSTIAN, KA [1 ]
FOOR, F [1 ]
机构
[1] MERCK & CO INC, RES LABS, MICROBIOL CHEMOTHERAPEUT & MOLEC GENET, RAHWAY, NJ 07065 USA
来源
JOURNAL OF GENERAL MICROBIOLOGY | 1993年 / 139卷
关键词
D O I
10.1099/00221287-139-12-2973
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
The immunosuppressants FK506 and cyclosporin A (CsA) bound to their receptors, FKBP12 or cyclophilin, inhibit the Ca2+/calmodulin-dependent protein phosphatase, calcineurin, preventing T cell activation or, in yeast, recovery from a-mating factor arrest. Vegetative growth of yeast does not require calcineurin, and in strains sensitive to FK506 or CsA, growth is inhibited by concentrations of drug much higher than those required to inhibit T cell activation or recovery from mating factor arrest. We now describe the isolation of a mutant of Saccharomyces cerevisiae which is 100-1000-fold more sensitive to the growth inhibitory properties of these drugs. The mutation (fks1) also confers a slow growth phenotype which is partially suppressed by exogenously added Ca2+ and exacerbated by EGTA. Simultaneous disruption of the two genes (CNA1 and CNA2) encoding the alternative forms of the catalytic A subunit of calcineurin, or of the gene (CNB1) encoding the regulatory B subunit, is lethal in an fks1 mutant. Disruption of the gene encoding FKBP12 (FKB1) or the major, cytosolic cyclophilin (CPH1) in fks1 cells results in the loss of hypersensitivity to the relevant drug. Overexpression of CNA1 or CNA2, in conjunction with CNB1, results in a significant decrease in hypersensitivity to FK506 and CsA. The results show that the hypersensitivity of the fks1 mutant is due to the inhibition of calcineurin phosphatase activity by the receptor-drug complexes. The growth dependence of the mutant on the Ca2+/calcineurin signal pathway provides an important tool for studying in yeast certain aspects of immune suppression by these drugs.
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页码:2973 / 2984
页数:12
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