A novel small molecule regulator of guanine nucleotide exchange activity of the ADP-ribosylation factor and golgi membrane trafficking.
An image-based phenotypic screen was developed to identify small molecule regulators of intracellular traffic. Using this screen we found that AG1478, a previously known inhibitor of epidermal growth factor receptor, had epidermal growth factor receptor-independent activity in inducing the disassembly of the Golgi in human cells. Similar to brefeldin A (BFA), a known disrupter of the Golgi, AG1478 inhibits the activity of small GTPase ADP-ribosylation factor. Unlike BFA, AG1478 exhibits low cytotoxicity and selectively targets the cis-Golgi without affecting endosomal compartment. We show that AG1478 inhibits GBF1, a large nucleotide exchange factor for the ADP-ribosylation factor, in a Sec7 domain-dependent manner and mimics the phenotype of a GBF1 mutant that has an inactive mutation. The treatment with AG1478 leads to the recruitment of GBF1 to the vesicular-tubular clusters adjacent to the endoplasmic reticulum exit sites, a step only transiently observed previously in the presence of BFA. We propose that the treatment with AG1478 delineates a membrane trafficking intermediate step that depends upon the Sec7 domain.
Pan
Heling
H
State Key Laboratory of Bio-organic & Natural Products Chemistry, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Shanghai 200032, China.
Yu
Jia
J
Zhang
Lihong
L
Carpenter
Anne
A
Zhu
Hong
H
Li
Li
L
Ma
Dawei
D
Yuan
Junying
J
D001693:Biological Transport, Active / Q000187:drug effects / Q000235:genetics; D020126:Brefeldin A / Q000494:pharmacology; D002460:Cell Line; D006056:Golgi Apparatus / Q000378:metabolism*; D020662:Guanine Nucleotide Exchange Factors / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007425:Intracellular Membranes / Q000378:metabolism*; D009154:Mutation; D017434:Protein Structure, Tertiary / Q000235:genetics; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology*; D011799:Quinazolines; D020032:Tyrphostins / Q000494:pharmacology*
C120445:GBF1 protein, human; D020662:Guanine Nucleotide Exchange Factors; D011500:Protein Synthesis Inhibitors; D011799:Quinazolines; D020032:Tyrphostins; C101044:RTKI cpd; D020126:Brefeldin A
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