{"doi":"10.1101/2020.05.22.111153","title":"Reciprocal regulation of vacuolar calcium transport and V-ATPase activity, and the effects of Phosphatidylinositol 3,5-bisphosphate","abstract":"ABSTRACT Yeast vacuoles are acidified by the V-ATPase, a protein complex comprised of the membrane embedded V O complex and the soluble cytoplasmic V 1 complex. The assembly of the V 1 -V O holoenzyme is required for the transfer of H + into the vacuole lumen for acidification. The assembly of the V 1 -V O holoenzyme is stabilized by the lipid phosphatidylinositol 3,5-bisphospate (PI(3,5)P 2 ) made by the PI3P 5-kinase Fab1/PIKfyve. The absence of PI(3,5)P 2 leads to the dissociation of the V 1 complex from the membrane. Separately, PI(3,5)P 2 has been shown to modulate Ca 2+ transport across the vacuole membrane during fission and fusion. Here we examined whether the regulation of H + and Ca 2+ by PI(3,5)P 2 are interdependent. We show that modulating extraluminal Ca 2+ concentrations inhibit V-ATPase activity. As extraluminal CaCl 2 levels are raised, the activity of H + pumping is reduced. Conversely, chelating free Ca 2+ with EGTA stimulated vacuole acidification. Not only did Ca 2+ levels affect H + translocation, we also show that blocking V-ATPase activity inhibited Ca 2+ transport into the vacuole lumen. Together, these data illustrate that Ca 2+ transport and V-ATPase regulation are interconnected through the modulation of vacuolar lipid profiles. Summary Statement Here we show that Ca 2+ and H + transport across the vacuole membrane is reciprocally regulated and that it is linked to the production of Phosphatidylinositol 3,5-bisphoshpate.","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":2020,"id":123682,"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":3,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9566,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2020-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":369256,"name":"David A. Rivera‐Kohr","orcid":"0000-0001-7199-3490","position":1,"is_corresponding":false},{"id":369255,"name":"Chi Zhang","orcid":"0000-0002-7735-5614","position":2,"is_corresponding":false},{"id":369254,"name":"Katherine D. Sullivan","orcid":"0000-0002-4898-5641","position":3,"is_corresponding":false},{"id":369257,"name":"Annie Guo","orcid":"0000-0003-4635-1183","position":4,"is_corresponding":false},{"id":369259,"name":"Rutilio A. Fratti","orcid":"0000-0001-9109-6666","position":5,"is_corresponding":false},{"id":369253,"name":"Gregory E. Miner","orcid":"0000-0002-1124-4217","position":0,"is_corresponding":true}],"reference_count":55,"raw_metadata":null,"created_at":"2026-07-18T23:15:03.403566Z","pmid":null,"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":[]}