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Journal of Cell Science, Vol 103, Issue 3 773-784, Copyright © 1992 by Company of Biologists
JOURNAL ARTICLES |
EB Cluett and WJ Brown
Section of Biochemistry, Molecular and Cell Biology, Cornell University, Ithaca, NY 14853.
We have investigated the nature of the component(s) responsible for holding the cisternal membranes of the Golgi complex into a stacked unit. Isolated Golgi complexes were treated with a variety of agents to induce the separation of intact Golgi stacks into single cisternal elements, i.e. "unstacking", and the effects were analyzed and quantitated by electron microscopy. In control experiments, isolated, intact Golgi stacks were stable at 4 degrees C and 20 degrees C for > or = 1 h; however, some unstacking occurred at 32 degrees C. Treatment of intact Golgi stacks with a variety of proteolytic enzymes resulted in a time- and dose-dependent unstacking of the cisternae, although stacks were resistant to various other proteases. Following liberation from the stack, single cisternae remained flattened with dilated rims. The integrity of intact Golgi stacks was unaffected by treatment with various concentrations and combinations of monovalent and divalent cations, or chelators of divalent cations. Electron microscopic observations of tannic acid- or negatively stained Golgi complexes, revealed the presence of highly structured, intercisternal "bridges". When seen within intact Golgi complexes, these bridges were only consistently found between closely apposed cisternae and were not observed on dilated rims or secretory vesicles. These bridges, on both intact stacks and physically disrupted cisternae, were rectangular, being approximately 8.5 nm in width, approximately 11 nm in height. Treatment with proteases under conditions that resulted in the with proteases under conditions that resulted in the unstacking of intact complexes also removed these bridge structures. These data show that proteinaceous components are responsible for holding Golgi cisternae together into a cohesive, stacked unit, and identify a candidate bridge structure that could serve this purpose.
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