Arv1 regulates PM and ER membrane structure and homeostasis but is dispensable for intracellular sterol transport.
Arv1 regulates PM and ER membrane structure and homeostasis but is dispensable for intracellular sterol transport.
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DOI：
10.1111/tra.12082
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发表时间：
2013-08
期刊：
影响因子：
--
通讯作者：
Menon AK
中科院分区：
文献类型：
--
作者：
Georgiev AG;Johansen J;Ramanathan VD;Sere YY;Beh CT;Menon AK
泛真核内质网(ER)膜蛋白Arv1被认为在细胞内固醇转运中发挥作用。我们通过比较野生型和Arv1缺陷型酿酒酵母中的甾醇运输来测试这一提议。我们使用荧光显微镜跟踪外源供应的脱氢麦角甾醇(DHE)从质膜(PM)到内质网和脂滴的逆行运动，并使用高效液相色谱平行定量DHE酯的运输耦合形成。利用代谢标记和亚细胞分级来分析麦角甾醇从ER到PM的顺行转运我们报道，内质网和质膜之间的类固醇运输不受Arv1缺乏的影响。相反，我们的结果表明，野生型和Arv1型Δ细胞在内质网形态和PM脂双层的组织结构上存在差异，这表明Arv1在膜动态平衡中具有明显的作用。在Arv1Δ细胞中，影响单个C末端跨膜结构域蛋白的特异性缺陷提示Arv1可能调节参与膜内稳态的尾部锚定蛋白的膜插入。
The pan-eukaryotic endoplasmic reticulum (ER) membrane protein Arv1 has been suggested to play a role in intracellular sterol transport. We tested this proposal by comparing sterol traffic in wild-type and Arv1-deficient Saccharomyces cerevisiae. We used fluorescence microscopy to track the retrograde movement of exogenously supplied dehydroergosterol (DHE) from the plasma membrane (PM) to the ER and lipid droplets and high performance liquid chromatography to quantify, in parallel, the transport-coupled formation of DHE esters. Metabolic labeling and subcellular fractionation were used to assay anterograde transport of ergosterol from the ER to the PM. We report that sterol transport between the ER and PM is unaffected by Arv1 deficiency. Instead, our results indicate differences in ER morphology and the organization of the PM lipid bilayer between wild-type and arv1Δ cells suggesting a distinct role for Arv1 in membrane homeostasis. In arv1Δ cells, specific defects affecting single C-terminal transmembrane domain proteins suggest that Arv1 might regulate membrane insertion of tail-anchored proteins involved in membrane homoeostasis.
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