Commonly used trafficking blocks disrupt ARF1 activation and the localization and function of specific Golgi proteins

Catherine E. Gilbert, Elizabeth Sztul, Carolyn E Machamer, Jennifer Lippincott-Schwartz

Research output: Contribution to journalArticle

Abstract

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.

Original languageEnglish (US)
Pages (from-to)937-947
Number of pages11
JournalMolecular Biology of the Cell
Volume29
Issue number8
DOIs
StatePublished - Apr 15 2018

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ADP-Ribosylation Factors
Brefeldin A
Guanine Nucleotide Exchange Factors
Proteins
Temperature
Monomeric GTP-Binding Proteins
Secretory Pathway
GTP Phosphohydrolases
Protein Transport
Guanosine Triphosphate
Membranes
ADP-ribosylation factor related proteins

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

Cite this

Commonly used trafficking blocks disrupt ARF1 activation and the localization and function of specific Golgi proteins. / Gilbert, Catherine E.; Sztul, Elizabeth; Machamer, Carolyn E; Lippincott-Schwartz, Jennifer.

In: Molecular Biology of the Cell, Vol. 29, No. 8, 15.04.2018, p. 937-947.

Research output: Contribution to journalArticle

Gilbert, Catherine E. ; Sztul, Elizabeth ; Machamer, Carolyn E ; Lippincott-Schwartz, Jennifer. / Commonly used trafficking blocks disrupt ARF1 activation and the localization and function of specific Golgi proteins. In: Molecular Biology of the Cell. 2018 ; Vol. 29, No. 8. pp. 937-947.
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