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Transport Through the Secretory Pathway: Observations of Cargo and Peripheral Coat Proteins

Published online by Cambridge University Press:  02 July 2020

John F. Presley
Affiliation:
Cell Biology and Metabolism Branch, Bldg 18T, NICHD, National Institutes of Health, Bethesda, MD20892.
Nelson B. Cole
Affiliation:
Cell Biology and Metabolism Branch, Bldg 18T, NICHD, National Institutes of Health, Bethesda, MD20892.
Jennifer Lippincott-Schwartz
Affiliation:
Cell Biology and Metabolism Branch, Bldg 18T, NICHD, National Institutes of Health, Bethesda, MD20892.
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Extract

We have used green fluorescent protein (GFP) chimeras to examine the dynamics of the early secretory pathway and the role of the peripheral coat protein COP I. To describe the overall properties of ER to Golgi transport we used the temperature sensitive viral glycoprotein, ts045 VSVG, tagged with GFP at its cytoplasmic tail. VSVG-GFP retained the temperature sensitive phenotype of its parent: it reversibly misfolded and was retained in the ER at 40°C. Upon shift to 32°C it was rapidly exported from the ER, moving as a synchronous pool into the Golgi complex and then to the cell surface. Using time-lapse imaging of living cells expressing VSVG-GFP we found that the carriers for ER to Golgi traffic are tubulovesicular pre-Golgi intermediates that move centrosomally to the Golgi at speeds of>1 μM2/sec and then fuse with the cis face of the Golgi complex. These movements are dependant on microtubules and the dynein/dynactin complex.

Type
Dynamics of Cellular Membrane Traffic
Copyright
Copyright © Microscopy Society of America

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