Compensatory Internalization of Pma1 in V-ATPase Mutants in Saccharomyces cerevisiae Requires Calcium- and Glucose-Sensitive Phosphatases

Genetics: A Periodical Record of Investigations Bearing on Heredity and Variation

Swetha Devi Velivela^1¹ 

[1] Department of Biochemistry and Molecular Biology, State University of New York Upstate Medical University, Syracuse, New York 13210^1

关键词: acidification;    arrestin;    calcineurin;    endocytosis;    phosphatase;    proton pump;    synthetic lethality;    ubiquitination;    vacuole;   

DOI  :  10.1534/genetics.117.300594

学科分类：医学（综合）

来源: Genetics Society of America

【 摘 要 】

Loss of V-ATPase activity in organelles, whether through V-ATPase inhibition or V-ATPase ( vma ) mutations, triggers a compensatory downregulation of the essential plasma membrane proton pump Pma1 in Saccharomyces cerevisiae . We have previously determined that the α-arrestin Rim8 and ubiquitin ligase Rsp5 are essential for Pma1 ubiquination and endocytosis in response to loss of V-ATPase activity. Here, we show that Pma1 endocytosis in V-ATPase mutants does not require Rim101 pathway components upstream and downstream of Rim8 , indicating that Rim8 is acting independently in Pma1 internalization. We find that two phosphatases, the calcium-responsive phosphatase calcineurin and the glucose-sensitive phosphatase Glc7 ( PP1 ), and one of the Glc7 regulatory subunits Reg1 , exhibit negative synthetic genetic interactions with vma mutants, and demonstrate that both phosphatases are essential for ubiquitination and endocytic downregulation of Pma1 in these mutants. Although both acute and chronic loss of V-ATPase activity trigger the internalization of ∼50% of surface Pma1 , a comparable reduction in Pma1 expression in a pma1 -007 mutant neither compensates for loss of V-ATPase activity nor stops further Pma1 endocytosis. The results indicate that the cell surface level of Pma1 is not directly sensed and that internalized Pma1 may play a role in compensating for loss of V-ATPase-dependent acidification. Taken together, these results provide new insights into cross talk between two major proton pumps central to cellular pH control.

【 授权许可】

CC BY