Evaluation of sterol transport from the endoplasmic reticulum to mitochondria using mitochondrially targeted bacterial sterol acyltransferase in Saccharomyces cerevisiae. Issue 10 (3rd October 2015)

• Record Type: Journal Article
• Authors: Tian, Siqi; Ohta, Akinori; Horiuchi, Hiroyuki; Fukuda, Ryouichi
• Is Part Of: Bioscience, biotechnology, and biochemistry. Volume 79:Issue 10(2015)
• Page Start: 1608
• Page End: 1614
• Publication Date: 2015-10-03
• Subjects: Saccharomyces cerevisiae -- sterol -- mitochondria -- endoplasmic reticulum -- transport; Biotechnology -- Periodicals; Biochemistry -- Periodicals; 660.6
• DOI: 10.1080/09168451.2015.1058702
• Languages: English
• ISSNs: 0916-8451

Abstract

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. Graphical abstract: : A system to evaluate sterol transport from the ER to mitochondria by measuring sterol esterification catalyzed by a mitochondrially targeted acyltransferase was constructed.