{"doi":"10.1016/j.jbc.2022.102672","title":"The interdependent transport of yeast vacuole Ca2+ and H+ and the role of phosphatidylinositol 3,5-bisphosphate","abstract":"Yeast vacuoles are acidified by the v-type H + -ATPase (V-ATPase) that is comprised of the membrane embedded V O complex and the soluble cytoplasmic V 1 complex. The assembly of the V 1 -V O holoenzyme on the vacuole is stabilized in part through interactions between the V O a-subunit ortholog Vph1 and the lipid phosphatidylinositol 3,5-bisphosphate (PI(3,5)P 2 ). PI(3,5)P 2 also affects vacuolar Ca 2+ release through the channel Yvc1 and uptake through the Ca 2+ pump Pmc1. Here, we asked if H + and Ca 2+ transport activities were connected through PI(3,5)P 2 . We found that overproduction of PI(3,5)P 2 by the hyperactive fab1 T2250A mutant augmented vacuole acidification, whereas the kinase-inactive fab1 EEE mutant attenuated the formation of a H + gradient. Separately, we tested the effects of excess Ca 2+ on vacuole acidification. Adding micromolar Ca 2+ blocked vacuole acidification, whereas chelating Ca 2+ accelerated acidification. The effect of adding Ca 2+ on acidification was eliminated when the Ca 2+ /H + antiporter Vcx1 was absent, indicating that the vacuolar H + gradient can collapse during Ca 2+ stress through Vcx1 activity. This, however, was independent of PI(3,5)P 2 , suggesting that PI(3,5)P 2 plays a role in submicromolar Ca 2+ flux but not under Ca 2+ shock. To see if the link between Ca 2+ and H + transport was bidirectional, we examined Ca 2+ transport when vacuole acidification was inhibited. We found that Ca 2+ transport was inhibited by halting V-ATPase activity with Bafilomycin or neutralizing vacuolar pH with chloroquine. Together, these data show that Ca 2+ transport and V-ATPase efficacy are connected but not necessarily through PI(3,5)P 2 .","journal":"Journal of Biological Chemistry","year":2022,"id":263664,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":13,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9479,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2022-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":690916,"name":"Yilin Feng","orcid":"0000-0002-9927-5658","position":1,"is_corresponding":false},{"id":565201,"name":"Adam Balutowski","orcid":null,"position":2,"is_corresponding":false},{"id":369253,"name":"Gregory E. Miner","orcid":"0000-0002-1124-4217","position":3,"is_corresponding":false},{"id":369256,"name":"David A. Rivera‐Kohr","orcid":"0000-0001-7199-3490","position":4,"is_corresponding":false},{"id":565200,"name":"Michael R. Hrabak","orcid":null,"position":5,"is_corresponding":false},{"id":369254,"name":"Katherine D. Sullivan","orcid":"0000-0002-4898-5641","position":6,"is_corresponding":false},{"id":369257,"name":"Annie Guo","orcid":"0000-0003-4635-1183","position":7,"is_corresponding":false},{"id":463213,"name":"Jorge D. Calderin","orcid":"0000-0002-9462-6180","position":8,"is_corresponding":false},{"id":369259,"name":"Rutilio A. Fratti","orcid":"0000-0001-9109-6666","position":9,"is_corresponding":false},{"id":920706,"name":"Chi Zhang","orcid":"0000-0002-4522-0598","position":0,"is_corresponding":true}],"reference_count":91,"raw_metadata":null,"created_at":"2026-07-19T00:26:33.413850Z","pmid":"36334632","pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}