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Papers In Press, published online ahead of print March 2, 2004
Biozentrum, University of Basel, Basel, CH 4056
Corresponding Author: Martin.Spiess{at}unibas.ch
The formation of secretory granules and regulated secretion are generally assumed to occur only in specialized endocrine, neuronal, or exocrine cells. We discovered that regulated secretory proteins such as the hormone precursors pro-vasopressin, pro-oxytocin, and pro-opiomelanocortin, as well as the granins secretogranin II and chromogranin B, but not the constitutive secretory protein
J. Biol. Chem, 10.1074/jbc.M310613200
Submitted on September 25, 2003
Revised on February 18, 2004
Accepted on March 2, 2004
Expression of regulated secretory proteins is sufficient to generate granule-like structures in constitutively secreting cells
1-protease inhibitor, accumulate in granular structures at the Golgi and in the cell periphery in transfected COS-1 fibroblast cells. The accumulations were observed in 30–70% of the transfected cells expressing the prohormones and for virtually all cells expressing the granins. Similar structures were also generated in other cell lines believed to be lacking a regulated secretory pathway. The accumulations resembled secretory granules morphologically in immunofluorescence and electron microscopy. They were devoid of markers of the endoplasmic reticulum, endosomes, and lysosomes, but in part stained positive for the trans-Golgi network marker TGN46, consistent with their formation at the trans-Golgi network. When different regulated proteins were coexpressed, they were frequently found in the same granules, whereas a1-protease inhibitor could not be detected in accumulations formed by secretogranin II, demonstrating segregation of regulated from constitutive secretory proteins. In pulse-chase experiments, significant intracellular storage of secretogranin II and chromogranin B was observed, and secretion of retained secretogranin II was stimulated with the calcium ionophore A23187. The results suggest that expression of regulated cargo proteins is sufficient to generate structures which resemble secretory granules in the background of constitutively secreting cells, supporting earlier proposals on the mechanism of granule formation.
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