Journal
JOURNAL OF NEUROCHEMISTRY
Volume 116, Issue 6, Pages 1122-1137Publisher
WILEY
DOI: 10.1111/j.1471-4159.2010.07167.x
Keywords
ADP-ribosylation factor 6; dystrophin; gephyrin; PDZ domain; post-synaptic density
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Funding
- Ministry of Education, Science, Sports, Culture, and Technology of Japan [2165049, 22300114]
- Kitasato University [2009-A01]
- Grants-in-Aid for Scientific Research [21500347, 22300114, 23659101, 19100005] Funding Source: KAKEN
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P>SynArfGEF, also known as BRAG3 or IQSEC3, is a member of the brefeldin A-resistant Arf-GEF/IQSEC family and was originally identified by screening for mRNA species associated with the post-synaptic density fraction. In this study, we demonstrate that synArfGEF activates Arf6, using Arf pull down and transferrin incorporation assays. Immunohistochemical analysis reveals that synArfGEF is present in somata and dendrites as puncta in close association with inhibitory synapses, whereas immunoelectron microscopic analysis reveals that synArfGEF localizes preferentially at post-synaptic specializations of symmetric synapses. Using yeast two-hybrid and pull down assays, we show that synArfGEF is able to bind utrophin/dystrophin and S-SCAM/MAGI-2 scaffolding proteins that localize at inhibitory synapses. Double immunostaining reveals that synArfGEF co-localizes with dystrophin and S-SCAM in cultured hippocampal neurons and cerebellar cortex, respectively. Both beta-dystroglycan and S-SCAM were immunoprecipitated from brain lysates using anti-synArfGEF IgG. Taken together, these findings suggest that synArfGEF functions as a novel regulator of Arf6 at inhibitory synapses and associates with the dystrophin-associated glycoprotein complex and S-SCAM.
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