title: SP1 regulates a human SNAP-25 gene expression.
abstract: The synaptosomal-associated protein of 25 kDa (SNAP-25) is a pre-synaptic plasma membrane protein. SNAP-25 plays an important role in synaptic vesicle membrane docking and fusion, which is involved in the regulation of neurotransmitter release. SNAP-25 has been implicated in the pathogenesis of neuropsychiatric disorders including Schizophrenia, attention-deficit hyperactivity disorder and Alzheimer's disease. We cloned a 1584 bp segment of the 5' flanking region of the human SNAP-25 gene. A series of nested deletions of the 5' flanking region fragment were subcloned into the pGL3-basic luciferase reporter plasmid. N2A cells were transfected with the SNAP-25 promoter constructs and luciferase activity was measured as an indication of promoter activity. We identified a 188 bp fragment containing the transcription initiation site as the minimal region necessary for promoter activity. Several putative cis-acting elements including SP1, hypoxia inducible factor (HIF), cAMP-response element binding protein, T-cell factor/lymphocyte enhancer factor 1 (TCF/LEF1), AP1 and the signal transducer and activator of transcription-6 (STAT6) are found in the 5' flanking region of SNAP-25 gene. Transcriptional activation and gel shift assays showed that the human SNAP-25 gene promoter contains functional SP1 response elements. Over-expression of SP1 increased SNAP-25 gene expression and inhibition of SP1-mediated transcriptional activation reduced SNAP-25 gene expression. These results suggest that SP1 plays an important role in regulation of the human SNAP-25 gene expression.
authors.affiliation: Department of Psychiatry, Brain Research Center, The University of British Columbia, Vancouver, British Columbia, Canada.
authors.forename: Fang
authors.initials: F
authors.lastname: Cai
authors.forename: Bin
authors.initials: B
authors.lastname: Chen
authors.forename: Weihui
authors.initials: W
authors.lastname: Zhou
authors.forename: Odysseus
authors.initials: O
authors.lastname: Zis
authors.forename: Shengchun
authors.initials: S
authors.lastname: Liu
authors.forename: Robert A
authors.initials: RA
authors.lastname: Holt
authors.forename: William G
authors.initials: WG
authors.lastname: Honer
authors.forename: Weihong
authors.initials: W
authors.lastname: Song
mesh_terms: D001483:Base Sequence; D001665:Binding Sites; D002461:Cell Line, Transformed; D024202:Electrophoretic Mobility Shift Assay / Q000379:methods; D015870:Gene Expression / Q000235:genetics*; D005786:Gene Expression Regulation / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D006801:Humans; D008926:Plicamycin / Q000031:analogs & derivatives / Q000494:pharmacology; D011401:Promoter Regions, Genetic / Q000502:physiology; D012045:Regulatory Sequences, Nucleic Acid; D017384:Sequence Deletion / Q000502:physiology; D016329:Sp1 Transcription Factor / Q000502:physiology*; D050825:Synaptosomal-Associated Protein 25 / Q000235:genetics / Q000378:metabolism*; D024363:Transcription Initiation Site / Q000502:physiology; D014158:Transcription, Genetic; D014162:Transfection / Q000379:methods
chemical_list: C494208:SNAP25 protein, human; D016329:Sp1 Transcription Factor; D050825:Synaptosomal-Associated Protein 25; C066851:mithramycin A; D008926:Plicamycin