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Synchronous intra-Golgi transport induces the release of Ca from the Golgi apparatus
Authors:Massimo Micaroni  Giuseppe Perinetti  Daniele Di Giandomenico  Alexander Spaar
Affiliation:a Department of Cell Biology and Oncology, Consorzio Mario Negri Sud, 66030 Santa Maria Imbaro (CH), Italy
b Department of Experimental and Diagnostic Medicine, Section of General Pathology, University of Ferrara, 44100 Ferrara, Italy
Abstract:The mechanisms of secretory transport through the Golgi apparatus remain an issue of debate. The precise functional importance of calcium ions (Ca2+) for intra-Golgi transport has also been poorly studied. Here, using different approaches to measure free Ca2+ concentrations in the cell cytosol ([Ca2+]cyt) and inside the lumen of the Golgi apparatus ([Ca2+]GA), we have revealed transient increases in [Ca2+]cyt during the late phase of intra-Golgi transport that are concomitant with a decline in the maximal [Ca2+]GA restoration ability. Thus, this redistribution of Ca2+ from the Golgi apparatus into the cytosol during the movement of cargo through the Golgi apparatus appears to have a role in intra-Golgi transport, and mainly in the late Ca2+-dependent phase of SNARE-regulated fusion of Golgi compartments.
Keywords:ARF, ADP-ribosylation factor   [Ca2+]cyt, free calcium ion concentration in the cell cytosol   ([Ca2+]ER, free calcium ion concentration in the endoplasmic reticulum   [Ca2+]GA, free calcium ion concentration in the Golgi apparatus   CHX, cycloheximide   COPI, coat protein I   ER, endoplasmic reticulum   Fluo-3-AM, acetoxymethyl ester form of the Ca2+-sensitive dye Fluo-3   FRET, fluorescence resonance energy transfer   FURA-2-AM, acetoxymethyl ester form of the Ca2+-sensitive dye FURA-2   GA, Golgi apparatus   GT, galactosyl transferase   HBSS, HEPES-buffered salt solution   HFs, human fibroblasts   PC-I, procollagen I   SNARE, soluble N-ethylmaleimide-sensitive factor attachment protein receptors   ST-Aeq, sialyl transferase-aequorin   VSVG, the ts045 temperature-sensitive variant of the G protein of vesicular stomatitis virus.
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