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© The Rockefeller University Press, 0021-9525/2000//1489 $5.00
The Journal of Cell Biology, Volume 150, Number 6, , 2000 1489-1498

Calcium Regulates the Association between Mitochondria and a Smooth Subdomain of the Endoplasmic Reticulum

^a Department of Pathology and Cell Biology, University of Montreal, Montreal, Quebec, Canada H3C 3J7
^b Department of Physiology, University of Montreal, Montreal, Quebec, Canada H3C 3J7
Département de pathologie et biologie cellulaire, Université de Montréal, C. P. 6128, succursale A, Montréal, Québec, Canada H3C 3J7.(514) 343-2459(514) 343-6291

Association between the ER and mitochondria has long been observed, and the formation of close contacts between ER and mitochondria is necessary for the ER-mediated sequestration of cytosolic calcium by mitochondria. Autocrine motility factor receptor (AMF-R) is a marker for a smooth subdomain of the ER, shown here by confocal microscopy to be distinct from, yet closely associated with the calnexin- or calreticulin-labeled ER. By EM, smooth ER AMF-R tubules exhibit direct interactions with mitochondria, identifying them as a mitochondria-associated smooth ER subdomain. In digitonin-permeabilized MDCK cells, the addition of rat liver cytosol stimulates the dissociation of smooth ER and mitochondria under conditions of low calcium. Using BAPTA chelators of various affinities and CaEGTA buffers of defined free Ca²⁺ concentrations and quantitative confocal microscopy, we show that free calcium concentrations <100 nM favor dissociation, whereas those >1 µM favor close association between these two organelles. Therefore, we describe a cellular mechanism that facilitates the close association of this smooth ER subdomain and mitochondria when cytosolic free calcium rises above physiological levels.

Key Words: free calcium • autocrine motility factor receptor • AMF-R tubule • cytosol • semipermeabilized cell assay

© 2000 The Rockefeller University Press

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