Participation of syntaxin 1A in membrane trafficking involving neurite elongation and membrane expansion.
Syntaxin 1A has been implicated to play an important role in neurotransmitter release by regulating synaptic vesicle fusion. The protein is also suggested to be required for other types of membrane fusion such as cellularization during embryonic development. In the current study, we overexpressed syntaxin 1A, SNAP-25b, and VAMP-2 in PC12 cells using recombinant adenoviruses, and determined their effects on membrane trafficking involving neurite outgrowth. It was found that overexpression of syntaxin 1A inhibited NGF-induced neurite extension, and the expressed syntaxin was localized to the plasma membrane, intracellular membranes, and the neurite tips. SNAP-25 overexpression slightly enhanced neurite elongation, whereas no significant changes in neurite growth was observed in VAMP-overexpressing cells. The effect of syntaxin 1A in general membrane trafficking was further studied by transient transfection of non-neuronal cells. Syntaxin 1A expression in HEK 239 and NIH3T3-L1 caused the cells to lose their normal morphology, leading to round and smaller cells. Deletion of the C-terminal sequence containing the H3 helix and the membrane anchoring domains of syntaxin abolished its ability to induce cell morphology changes, whereas removal of the N-terminal 1-170 amino acid sequence did not affect this activity. These findings suggest that in addition to its well documented role in synaptic vesicle fusion, syntaxin may function in other non-synaptic membrane trafficking such as neurite outgrowth and membrane expansion.
Zhou
Q
Q
Department of Pathology, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 73190, USA.
Xiao
J
J
Liu
Y
Y
D016475:3T3 Cells; D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D000954:Antigens, Surface / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D002462:Cell Membrane / Q000502:physiology; D015151:Immunoblotting; D007150:Immunohistochemistry; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D016501:Neurites / Q000502:physiology*; D009474:Neurons / Q000502:physiology* / Q000648:ultrastructure; D016716:PC12 Cells; D050683:R-SNARE Proteins; D051381:Rats; D050825:Synaptosomal-Associated Protein 25; D050827:Syntaxin 1; D014162:Transfection
D000954:Antigens, Surface; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; D050683:R-SNARE Proteins; C494209:Snap25 protein, mouse; C494210:Snap25 protein, rat; C494214:Stx1a protein, mouse; C494213:Stx1a protein, rat; D050825:Synaptosomal-Associated Protein 25; D050827:Syntaxin 1