The yeast mutant vps5Δ affected in the recycling of Golgi membrane proteins displays an enhanced vacuolar Mg2+/H+ exchange activity

被引:16
作者
Borrelly, G [1 ]
Boyer, JC [1 ]
Touraine, B [1 ]
Szponarski, W [1 ]
Rambier, M [1 ]
Gibrat, R [1 ]
机构
[1] Univ Montpellier 2, Inst Natl Rech Agron, CNRS, UMR 5004,ENSA M, F-34060 Montpellier 1, France
关键词
D O I
10.1073/pnas.161215198
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Growth of the yeast vacuolar protein-sorting mutant vps5 Delta affected in the endosome-to-Golgi retromer complex was more sensitive to Mg2+-limiting conditions than was the growth of the wild-type (WT) strain. This sensitivity was enhanced at acidic pH. The vps5 Delta strain was also sensitive to Al3+, known to inhibit Mg2+ uptake in yeast cells. In contrast, it was found to be resistant to Ni2+ and CO2+ two cytotoxic analogs of Mg2+. Resistance to Ni2+ did not seem to result from the alteration of plasma-membrane transport properties because mutant and WT cells displayed similar Ni2+ uptake. After plasma-membrane permeabilization, intracellular Ni2+ uptake in vps5 Delta cells was 3-fold higher than in WT cells, which is consistent with the implication of the vacuole in the observed phenotypes. In reconstituted vacuolar vesicles prepared from vps5 Delta, the rates of H+ exchange with Ni2+, CO2+, and Mg2+ were increased (relative to WT) by 170%, 130%, and 50%, respectively. The rates of H+ exchange with Ca2+, Cd2+, and K+ were similar in both strains, as were alpha -mannosiclase and H+-ATPase activities, and SDS/PAGE patterns of vacuolar proteins. Among 14 other vacuolar protein-sorting mutants tested, only the 8 mutants affected in the recycling of trans-Golgi network membrane proteins shared the same Ni2+ resistance phenotype as vps5 Delta. It is proposed that a trans-Golgi network Mg2+/H+ exchanger, mislocalized to vps5 Delta vacuole, could be responsible for the phenotypes observed in vivo and in vitro.
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页码:9660 / 9665
页数:6
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