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Published online 5 November 2001. doi:10.1083/jcb.200103104
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© The Rockefeller University Press, 0021-9525/2001/11/543 $5.00
The Journal of Cell Biology, Volume 155, Number 4, November 12, 2001 543-556

Article

 Evidence that the entire Golgi apparatus cycles in interphase HeLa cells : sensitivity of Golgi matrix proteins to an ER exit block



Suzanne Miles1, Heather McManus1, Kimberly E. Forsten2 and Brian Storrie1

1 Department of Biochemistry, Virginia Tech, Blacksburg, VA 24061
2 Department of Chemical Engineering, Virginia Tech, Blacksburg, VA 24061

Address correspondence to Brian Storrie, Department of Biochemistry, Virginia Tech, Blacksburg, VA 24061. Tel.: (540) 231-6434. Fax: (540) 231-9070. E-mail: storrie{at}vt.edu

We tested whether the entire Golgi apparatus is a dynamic structure in interphase mammalian cells by assessing the response of 12 different Golgi region proteins to an endoplasmic reticulum (ER) exit block. The proteins chosen spanned the Golgi apparatus and included both Golgi glycosyltransferases and putative matrix proteins. Protein exit from ER was blocked either by microinjection of a GTP-restricted Sar1p mutant protein in the presence of a protein synthesis inhibitor, or by plasmid-encoded expression of the same dominant negative Sar1p. All Golgi region proteins examined lost juxtanuclear Golgi apparatus–like distribution as scored by conventional and confocal fluorescence microscopy in response to an ER exit block, albeit with a differential dependence on Sar1p concentration. Redistribution of GalNAcT2 was more sensitive to low Sar1pdn concentrations than giantin or GM130. Redistribution was most rapid for p27, COPI, and p115. Giantin, GM130, and GalNAcT2 relocated with approximately equal kinetics. Distinct ER accumulation could be demonstrated for all integral membrane proteins. ER-accumulated Golgi region proteins were functional. Photobleaching experiments indicated that Golgi-to-ER protein cycling occurred in the absence of any ER exit block. We conclude that the entire Golgi apparatus is a dynamic structure and suggest that most, if not all, Golgi region–integral membrane proteins cycle through ER in interphase cells.

Key Words: Golgi apparatus; Sar1p; ER exit block; protein cycling; Golgi matrix


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