title: Evaluation of sterol transport from the endoplasmic reticulum to mitochondria using mitochondrially targeted bacterial sterol acyltransferase in Saccharomyces cerevisiae.
abstract: To elucidate the mechanism of interorganelle sterol transport, a system to evaluate sterol transport from the endoplasmic reticulum (ER) to the mitochondria was constructed. A bacterial glycerophospholipid: cholesterol acyltransferase fused with a mitochondria-targeting sequence and a membrane-spanning domain of the mitochondrial inner membrane protein Pet100 and enhanced green fluorescent protein was expressed in a Saccharomyces cerevisiae mutant deleted for ARE1 and ARE2 encoding acyl-CoA:sterol acyltransferases. Microscopic observation and subcellular fractionation suggested that this fusion protein, which was named mito-SatA-EGFP, was localized in the mitochondria. Steryl esters were synthesized in the mutant expressing mito-SatA-EGFP. This system will be applicable for evaluations of sterol transport from the ER to the mitochondria in yeast by examining sterol esterification in the mitochondria.
authors.affiliation: a Department of Biotechnology , The University of Tokyo , Tokyo , Japan.
authors.forename: Siqi
authors.initials: S
authors.lastname: Tian
authors.forename: Akinori
authors.initials: A
authors.lastname: Ohta
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Horiuchi
authors.forename: Ryouichi
authors.initials: R
authors.lastname: Fukuda
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D001692:Biological Transport; D002788:Cholesterol Esters / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D015966:Gene Expression Regulation, Fungal; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D008928:Mitochondria / Q000378:metabolism*; D018832:Molecular Chaperones / Q000235:genetics / Q000378:metabolism; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism; D002785:Sterol O-Acyltransferase / Q000172:deficiency / Q000235:genetics / Q000378:metabolism; D013261:Sterols / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D002788:Cholesterol Esters; D018832:Molecular Chaperones; D009687:Nuclear Proteins; C100954:PET100 protein, S cerevisiae; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; D013261:Sterols; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; C543386:ARE1 protein, S cerevisiae; C543388:ARE2 protein, S cerevisiae; D002785:Sterol O-Acyltransferase
keywords: Saccharomyces cerevisiae; endoplasmic reticulum; mitochondria; sterol; transport