Transcriptional activation of the neuronal peripherin-encoding gene depends on a G + C-rich element that binds Sp1 in vitro and in vivo.
Peripherin (Prph) is a type-III intermediate filament (IF) protein principally synthesized in peripheral nervous system neurons. We have previously shown that three regulatory elements, PER1, PER2 and PER3, in the first 98 bp of the Prph gene promoter, were sufficient to direct cell-type specific expression of a reporter gene [Desmarais et al., EMBO J. 11 (1992) 2971-2980]. Of these elements, PER1 was found to be important for cell-type specificity, but required the presence of other elements for transcriptional activity. Here, we show that PER3 is a stronger activator than PER2 and that it can stimulate cell-type-specific transcription when combined with PER1. We have characterized the G + C-rich PER3 element for its ability to bind trans-acting factors. Gel retardation and methylation interference (MI) assays show that PER3 binds transcription factor Sp1. In addition, an anti-Sp1 antibody recognizes the PER3 DNA-binding protein. A 3-bp mutation abrogating the capacity of PER3 to bind Sp1 in vitro completely abolished expression of the reporter gene construct containing only PER3 and PER1, while in a construct containing the first 256 bp of the Prph promoter, it led to an 80% decrease with respect to the control wild-type construct. Finally, by co-transfection of a Sp1-expressing plasmid, we show that Sp1 can stimulate transcription from a reporter gene containing the PER3 sequence. Together, these results indicate that interactions between Sp1 and the proteins binding PER1 are involved in the control of the Prph gene.
Ferrari
N
N
Département de Pathologie, Université de Montréal, Québec, Canada.
Desmarais
D
D
Royal
A
A
D001482:Base Composition; D001483:Base Sequence; D001665:Binding Sites; D004252:DNA Mutational Analysis; D005786:Gene Expression Regulation; D007381:Intermediate Filament Proteins / Q000096:biosynthesis / Q000235:genetics*; D008562:Membrane Glycoproteins; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins; D009474:Neurons / Q000502:physiology*; D064531:Peripherins; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D016329:Sp1 Transcription Factor / Q000378:metabolism*; D014158:Transcription, Genetic
D007381:Intermediate Filament Proteins; D008562:Membrane Glycoproteins; D009419:Nerve Tissue Proteins; D064531:Peripherins; D011993:Recombinant Fusion Proteins; D016329:Sp1 Transcription Factor