title: GLUT1 glucose transporter gene transcription is repressed by Sp3. Evidence for a regulatory role of Sp3 during myogenesis.
abstract: GLUT1 glucose transporters are highly expressed in proliferating and transformed cells as well as in tissues during fetal life. However, the mechanisms that regulate GLUT1 gene expression remain largely unknown. Here, we demonstrate that Sp3 proteins bind to the GLUT1 proximal promoter gene and inhibit transcriptional activity in muscle and non-muscle cells. Two different Sp3 translational products (110 and 74 kDa) derived from differential translational initiation were detected in nuclear extracts from myoblast cells, and both Sp3 protein species inhibited GLUT1 gene transcriptional activity. The inhibitory effect of Sp3 was dominant over the stimulatory effect of Sp1 on transcriptional activity of GLUT1 gene. Furthermore, abolition of Sp3 binding to the proximal promoter of GLUT1 gene completely blocked the response to Sp3. We provide evidence that the expression of Sp3 protein is subject to regulation in muscle cells and that this is likely to control GLUT1. Thus, Sp3 protein was up-regulated in the absence of changes in Sp1 early after the induction of IGF-II-dependent myogenesis. Furthermore, forced over-expression of MyoD caused an enhancement in the cellular Sp3/Sp1 ratio which was concomitant to a reduced GLUT1 expression. Later during myogenesis, Sp3 expression was substantial whereas Sp1 was markedly down-regulated. In summary, we provide direct evidence that the transcription factor Sp3 represses gene expression in non-muscle and muscle cells and this is likely to operate in fetal heart by binding to the GLUT1 gene promoter. This is the first description of a repressor of GLUT1 gene transcription. Furthermore, we propose that variations in the ratio of Sp3 versus Sp1 regulate GLUT1 promoter activity and this is crucial in the down-regulation of GLUT1 associated to myogenesis.
authors.affiliation: Departament de Bioquímica i Biologia Molecular, Facultat de Biologia, Universitat de Barcelona, Barcelona, Spain.
authors.forename: C
authors.initials: C
authors.lastname: Fandos
authors.forename: M
authors.initials: M
authors.lastname: Sánchez-Feutrie
authors.forename: T
authors.initials: T
authors.lastname: Santalucía
authors.forename: F
authors.initials: F
authors.lastname: Viñals
authors.forename: J
authors.initials: J
authors.lastname: Cadefau
authors.forename: A
authors.initials: A
authors.lastname: Gumà
authors.forename: R
authors.initials: R
authors.lastname: Cussó
authors.forename: P
authors.initials: P
authors.lastname: Kaliman
authors.forename: J
authors.initials: J
authors.lastname: Canicio
authors.forename: M
authors.initials: M
authors.lastname: Palacín
authors.forename: A
authors.initials: A
authors.lastname: Zorzano
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D004268:DNA-Binding Proteins / Q000302:isolation & purification / Q000378:metabolism*; D018507:Gene Expression Regulation, Developmental; D051272:Glucose Transporter Type 1; D007335:Insulin-Like Growth Factor II / Q000494:pharmacology; D009004:Monosaccharide Transport Proteins / Q000096:biosynthesis / Q000235:genetics*; D009121:Muscle Denervation; D018482:Muscle, Skeletal / Q000166:cytology*; D009687:Nuclear Proteins / Q000302:isolation & purification / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D020033:Protein Isoforms / Q000378:metabolism; D051381:Rats; D012097:Repressor Proteins / Q000302:isolation & purification / Q000378:metabolism*; D016329:Sp1 Transcription Factor / Q000378:metabolism; D051705:Sp3 Transcription Factor; D014018:Tissue Distribution; D014157:Transcription Factors / Q000302:isolation & purification / Q000378:metabolism*; D014158:Transcription, Genetic; D015854:Up-Regulation
chemical_list: D004268:DNA-Binding Proteins; D051272:Glucose Transporter Type 1; D009004:Monosaccharide Transport Proteins; D009687:Nuclear Proteins; D020033:Protein Isoforms; D012097:Repressor Proteins; C495679:Slc2a1 protein, rat; D016329:Sp1 Transcription Factor; C497123:Sp3 protein, rat; D014157:Transcription Factors; D051705:Sp3 Transcription Factor; D007335:Insulin-Like Growth Factor II