Ramakrishnan Srinivasan, Asady Beejan, Docampo Roberto
Center for Tropical and Emerging Global Diseases, University of Georgia, Athens, GA 30602, USA.
Department of Cellular Biology, University of Georgia, Athens, GA 30602, USA.
Pathogens. 2018 Mar 22;7(2):33. doi: 10.3390/pathogens7020033.
Membrane contact sites are regions of close apposition between two organelles, typically less than 30 nanometers apart, that facilitate transfer of biomolecules. The presence of contact sites has been demonstrated in yeast, plants, and mammalian cells. Here, we investigated the presence of such contact sites in . In mammalian cells, endoplasmic reticulum-mitochondria contact sites facilitate mitochondrial uptake of Ca released by the ER-located inositol 1,4,5-trisphosphate receptor (InsP₃R). However, the InsP₃R in trypanosomes localizes to acidocalcisomes, which serve as major Ca stores in these parasites. In this work, we have used super-resolution structured illumination microscopy and electron microscopy to identify membrane contact sites that exist between acidocalcisomes and mitochondria. Furthermore, we have confirmed the close association of these organelles using proximity ligation assays. Characterization of these contact sites may be a necessary starting point towards unraveling the role of Ca in regulating trypanosome bioenergetics.
膜接触位点是两个细胞器紧密相邻的区域,通常相距不到30纳米,有助于生物分子的转移。接触位点已在酵母、植物和哺乳动物细胞中得到证实。在此,我们研究了[具体生物]中此类接触位点的存在情况。在哺乳动物细胞中,内质网 - 线粒体接触位点促进位于内质网的肌醇1,4,5 - 三磷酸受体(InsP₃R)释放的Ca进入线粒体。然而,锥虫中的InsP₃R定位于酸性钙小体,酸性钙小体是这些寄生虫中主要的钙储存库。在这项工作中,我们使用超分辨率结构照明显微镜和电子显微镜来鉴定酸性钙小体和线粒体之间存在的膜接触位点。此外,我们使用邻近连接分析法证实了这些细胞器的紧密关联。对这些接触位点的表征可能是揭示Ca在调节锥虫生物能量学中作用的必要起点。