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Structure of the Golgi and Distribution of Reporter Molecules at 20°C Reveals the Complexity of the Exit Compartments

Ladinsky, Mark S. and Wu, Christine C. and McIntosh, Shane and McIntosh, J. Richard and Howell, Kathryn E. (2002) Structure of the Golgi and Distribution of Reporter Molecules at 20°C Reveals the Complexity of the Exit Compartments. Molecular Biology of the Cell, 13 (8). pp. 2810-2825. ISSN 1059-1524. PMCID PMC117944. doi:10.1091/mbc.01-12-0593.

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Incubating cells at 20°C blocks transport out of the Golgi complex and amplifies the exit compartments. We have used the 20°C block, followed by EM tomography and serial section reconstruction, to study the structure of Golgi exit sites in NRK cells. The dominant feature of Golgi structure in temperature-blocked cells is the presence of large bulging domains on the three trans-most cisternae. These domains extend laterally from the stack and are continuous with “cisternal” domains that maintain normal thickness and alignment with the other stacked Golgi cisternae. The bulging domains do not resemble the perpendicularly extending tubules associated with the trans-cisternae of control cells. Such tubules are completely absent in temperature-blocked cells. The three cisternae with bulging domains can be identified as trans by their association with specialized ER and the presence of clathrin-coated buds on the trans-most cisterna only. Immunogold labeling and immunoblots show a significant degradation of a medial- and a trans-Golgi marker with no evidence for their redistribution within the Golgi or to other organelles. These data suggest that exit from the Golgi occurs directly from three trans-cisternae and that specialized ER plays a significant role in trans-Golgi function.

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URLURL TypeDescription CentralArticle
Ladinsky, Mark S.0000-0002-1036-3513
Additional Information:© 2002 The American Society for Cell Biology. Submitted: 26 December 2001; Revised: 24 April 2002; Accepted: 1 May 2002. We thank Brad Marsh for critical reading of the manuscript and Mary Morphew, Eileen O'Toole, and David Mastronarde for helpful comments and suggestions through the course of the project. We also thank George Banting for the antibodies against the luminal domain of TGN38 (2F7) and Nicholas Gonatas for the antibodies against MG160. This project was funded by National Institutes of Health grant PO1-GM61306 to K.E.H. and J.R.M. and GM42629 to K.E.H.
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Issue or Number:8
PubMed Central ID:PMC117944
Record Number:CaltechAUTHORS:20191106-093627984
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Official Citation:Structure of the Golgi and Distribution of Reporter Molecules at 20°C Reveals the Complexity of the Exit Compartments Mark S. Ladinsky, Christine C. Wu, Shane McIntosh, J. Richard McIntosh, and Kathryn E. Howell Molecular Biology of the Cell 2002 13:8, 2810-2825
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:99685
Deposited By: Tony Diaz
Deposited On:06 Nov 2019 17:46
Last Modified:16 Nov 2021 17:48

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