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Commonly used trafficking blocks disrupt ARF1 activation and the localization and function of specific Golgi proteins
ADP-ribosylation factor (ARF) proteins are key regulators of the secretory pathway. ARF1, through interacting with its effectors, regulates protein trafficking by facilitating numerous events at the Golgi. One unique ARF1 effector is golgin-160, which promotes the trafficking of only a specific subs...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The American Society for Cell Biology
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5896932/ https://www.ncbi.nlm.nih.gov/pubmed/29467256 http://dx.doi.org/10.1091/mbc.E17-11-0622 |
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author | Gilbert, Catherine E. Sztul, Elizabeth Machamer, Carolyn E. |
author_facet | Gilbert, Catherine E. Sztul, Elizabeth Machamer, Carolyn E. |
author_sort | Gilbert, Catherine E. |
collection | PubMed |
description | ADP-ribosylation factor (ARF) proteins are key regulators of the secretory pathway. ARF1, through interacting with its effectors, regulates protein trafficking by facilitating numerous events at the Golgi. One unique ARF1 effector is golgin-160, which promotes the trafficking of only a specific subset of cargo proteins through the Golgi. While studying this role of golgin-160, we discovered that commonly used cold temperature blocks utilized to synchronize cargo trafficking (20 and 16°C) caused golgin-160 dispersal from Golgi membranes. Here, we show that the loss of golgin-160 localization correlates with a decrease in the levels of activated ARF1, and that golgin-160 dispersal can be prevented by expression of a GTP-locked ARF1 mutant. Overexpression of the ARF1 activator Golgi brefeldin A–resistant guanine nucleotide exchange factor 1 (GBF1) did not prevent golgin-160 dispersal, suggesting that GBF1 may be nonfunctional at lower temperatures. We further discovered that several other Golgi resident proteins had altered localization at lower temperatures, including proteins recruited by ARF-like GTPase 1 (ARL1), a small GTPase that also became dispersed in the cold. Although cold temperature blocks are useful for synchronizing cargo trafficking through the Golgi, our data indicate that caution must be taken when interpreting results from these assays. |
format | Online Article Text |
id | pubmed-5896932 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | The American Society for Cell Biology |
record_format | MEDLINE/PubMed |
spelling | pubmed-58969322018-06-30 Commonly used trafficking blocks disrupt ARF1 activation and the localization and function of specific Golgi proteins Gilbert, Catherine E. Sztul, Elizabeth Machamer, Carolyn E. Mol Biol Cell Articles ADP-ribosylation factor (ARF) proteins are key regulators of the secretory pathway. ARF1, through interacting with its effectors, regulates protein trafficking by facilitating numerous events at the Golgi. One unique ARF1 effector is golgin-160, which promotes the trafficking of only a specific subset of cargo proteins through the Golgi. While studying this role of golgin-160, we discovered that commonly used cold temperature blocks utilized to synchronize cargo trafficking (20 and 16°C) caused golgin-160 dispersal from Golgi membranes. Here, we show that the loss of golgin-160 localization correlates with a decrease in the levels of activated ARF1, and that golgin-160 dispersal can be prevented by expression of a GTP-locked ARF1 mutant. Overexpression of the ARF1 activator Golgi brefeldin A–resistant guanine nucleotide exchange factor 1 (GBF1) did not prevent golgin-160 dispersal, suggesting that GBF1 may be nonfunctional at lower temperatures. We further discovered that several other Golgi resident proteins had altered localization at lower temperatures, including proteins recruited by ARF-like GTPase 1 (ARL1), a small GTPase that also became dispersed in the cold. Although cold temperature blocks are useful for synchronizing cargo trafficking through the Golgi, our data indicate that caution must be taken when interpreting results from these assays. The American Society for Cell Biology 2018-04-15 /pmc/articles/PMC5896932/ /pubmed/29467256 http://dx.doi.org/10.1091/mbc.E17-11-0622 Text en © 2018 Gilbert et al. “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society for Cell Biology. http://creativecommons.org/licenses/by-nc-sa/3.0/ This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License. |
spellingShingle | Articles Gilbert, Catherine E. Sztul, Elizabeth Machamer, Carolyn E. Commonly used trafficking blocks disrupt ARF1 activation and the localization and function of specific Golgi proteins |
title | Commonly used trafficking blocks disrupt ARF1 activation and the localization and function of specific Golgi proteins |
title_full | Commonly used trafficking blocks disrupt ARF1 activation and the localization and function of specific Golgi proteins |
title_fullStr | Commonly used trafficking blocks disrupt ARF1 activation and the localization and function of specific Golgi proteins |
title_full_unstemmed | Commonly used trafficking blocks disrupt ARF1 activation and the localization and function of specific Golgi proteins |
title_short | Commonly used trafficking blocks disrupt ARF1 activation and the localization and function of specific Golgi proteins |
title_sort | commonly used trafficking blocks disrupt arf1 activation and the localization and function of specific golgi proteins |
topic | Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5896932/ https://www.ncbi.nlm.nih.gov/pubmed/29467256 http://dx.doi.org/10.1091/mbc.E17-11-0622 |
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