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PMID:21198641

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Citation

Fukaya, M, Kamata, A, Hara, Y, Tamaki, H, Katsumata, O, Ito, N, Takeda, S, Hata, Y, Suzuki, T, Watanabe, M, Harvey, RJ and Sakagami, H (2011) SynArfGEF is a guanine nucleotide exchange factor for Arf6 and localizes preferentially at post-synaptic specializations of inhibitory synapses. J. Neurochem. 116:1122-37

Abstract

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 β-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.

Links

PubMed Online version:10.1111/j.1471-4159.2010.07167.x

Keywords

ADP-Ribosylation Factors/genetics; ADP-Ribosylation Factors/metabolism; Adaptor Proteins, Signal Transducing; Animals; Brain/cytology; Carrier Proteins/metabolism; Cell Line, Transformed; Cercopithecus aethiops; Dystrophin/metabolism; Gene Expression Regulation/physiology; Guanine Nucleotide Exchange Factors/genetics; Guanine Nucleotide Exchange Factors/metabolism; Guanosine Triphosphate/metabolism; Guanylate Kinase; Humans; Immunoprecipitation/methods; Mice; Neurons/cytology; Neurons/metabolism; Protein Binding; Proteins/metabolism; Synapses/metabolism; Synapses/ultrastructure; Transfection/methods; Two-Hybrid System Techniques; Vesicular Inhibitory Amino Acid Transport Proteins/metabolism

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

MOUSE:IQEC3

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q9WVQ1

F

Seeded From UniProt

complete

MOUSE:IQEC3

located_in

GO:0045211: postsynaptic membrane

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:IQEC3

located_in

GO:0060077: inhibitory synapse

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:IQEC3

is_active_in

GO:0098982: GABA-ergic synapse

ECO:0005589: confocal microscopy evidence used in manual assertion

C

  • part_of:(UBERON:0002421)

Seeded From UniProt

complete

MOUSE:IQEC3

is_active_in

GO:0098982: GABA-ergic synapse

ECO:0005589: confocal microscopy evidence used in manual assertion

C

  • part_of:(UBERON:0002037)

Seeded From UniProt

complete

MOUSE:IQEC3

is_active_in

GO:0098982: GABA-ergic synapse

ECO:0006003: electron microscopy evidence used in manual assertion

C

  • part_of:(UBERON:0002421)

Seeded From UniProt

complete

MOUSE:IQEC3

is_active_in

GO:0098982: GABA-ergic synapse

ECO:0006003: electron microscopy evidence used in manual assertion

C

  • part_of:(UBERON:0002037)

Seeded From UniProt

complete

MOUSE:IQEC3

is_active_in

GO:0099629: postsynaptic specialization of symmetric synapse

ECO:0005589: confocal microscopy evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:IQEC3

is_active_in

GO:0099629: postsynaptic specialization of symmetric synapse

ECO:0005589: confocal microscopy evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:IQEC3

is_active_in

GO:0099629: postsynaptic specialization of symmetric synapse

ECO:0005589: confocal microscopy evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:IQEC3

is_active_in

GO:0099629: postsynaptic specialization of symmetric synapse

ECO:0006003: electron microscopy evidence used in manual assertion

C

  • part_of:(UBERON:0002264)

Seeded From UniProt

complete

MOUSE:IQEC3

is_active_in

GO:0099629: postsynaptic specialization of symmetric synapse

ECO:0006003: electron microscopy evidence used in manual assertion

C

  • part_of:(UBERON:0002037)

Seeded From UniProt

complete

MOUSE:IQEC3

is_active_in

GO:0099629: postsynaptic specialization of symmetric synapse

ECO:0006003: electron microscopy evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:IQEC3

is_active_in

GO:0099629: postsynaptic specialization of symmetric synapse

ECO:0006003: electron microscopy evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:IQEC3

is_active_in

GO:0099629: postsynaptic specialization of symmetric synapse

ECO:0006003: electron microscopy evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:IQEC3

is_active_in

GO:0098690: glycinergic synapse

ECO:0005589: confocal microscopy evidence used in manual assertion

C

  • part_of:(UBERON:0002240)

Seeded From UniProt

complete

MOUSE:IQEC3

is_active_in

GO:0098690: glycinergic synapse

ECO:0006003: electron microscopy evidence used in manual assertion

C

  • part_of:(UBERON:0002240)

Seeded From UniProt

complete

RAT:IQEC3

involved_in

GO:0051056: regulation of small GTPase mediated signal transduction

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

RAT:IQEC3

involved_in

GO:0090630: activation of GTPase activity

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

MOUSE:MAGI2

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q3TES0

F

Seeded From UniProt

complete


See also

References

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