title: The yeast vacuolar ABC transporter Ybt1p regulates membrane fusion through Ca2+ transport modulation.
abstract: Ybt1p is a class C ABC transporter (ATP-binding cassette transporter) that is localized to the vacuole of Saccharomyces cerevisiae. Although Ybt1p was originally identified as a bile acid transporter, it has also been found to function in other capacities, including the translocation of phosphatidylcholine to the vacuole lumen, and the regulation of Ca2+ homoeostasis. In the present study we found that deletion of YBT1 enhanced in vitro homotypic vacuole fusion by up to 50% relative to wild-type vacuoles. The increased vacuole fusion was not due to aberrant protein sorting of SNAREs (soluble N-ethylmaleimide-sensitive factor-attachment protein receptors) or recruitment of factors from the cytosol such as Ypt7p and the HOPS (homotypic fusion and vacuole protein sorting) tethering complex. In addition, ybt1Δ vacuoles displayed no observable differences in the formation of SNARE complexes, interactions between SNAREs and HOPS, or formation of vertex microdomains. However, the absence of Ybt1p caused significant changes in Ca2+ transport during fusion. One difference was the prolonged Ca2+ influx exhibited by ybt1Δ vacuoles at the start of the fusion reaction. We also observed a striking delay in SNARE-dependent Ca2+ efflux. As vacuole fusion can be inhibited by high Ca2+ concentrations, we suggest that the delayed efflux in ybt1Δ vacuoles leads to the enhanced SNARE function.
authors.affiliation: Department of Biochemistry, University of Illinois at Urbana-Champaign, Urbana, IL 61801, U.S.A.
authors.forename: Terry L
authors.initials: TL
authors.lastname: Sasser
authors.forename: Mark
authors.initials: M
authors.lastname: Padolina
authors.forename: Rutilio A
authors.initials: RA
authors.lastname: Fratti
mesh_terms: D018528:ATP-Binding Cassette Transporters / Q000172:deficiency / Q000502:physiology*; D002118:Calcium / Q000502:physiology*; D017353:Gene Deletion; D008561:Membrane Fusion / Q000502:physiology*; D021381:Protein Transport / Q000502:physiology; D050600:SNARE Proteins / Q000502:physiology; D012441:Saccharomyces cerevisiae / Q000737:chemistry / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000502:physiology*; D015854:Up-Regulation / Q000235:genetics; D014617:Vacuoles / Q000737:chemistry / Q000502:physiology*
chemical_list: D018528:ATP-Binding Cassette Transporters; D050600:SNARE Proteins; D029701:Saccharomyces cerevisiae Proteins; C106609:YBT1 protein, S cerevisiae; D002118:Calcium