An upstream stimulatory factor (USF) binding motif is critical for rat preprotachykinin-A promoter activity in PC12 cells.
We demonstrate the presence of a functional E box motif in the proximal rat preprotachykinin-A (rPPT) promoter. This element (spanning nucleotides -67 to -47) exhibits the sequence 5'-CACGTG-3' which is recognized and bound by the basic helix-loop-helix family of regulatory proteins. We also show that at least one of the factors bound to this rPPT promoter element in both HeLa and PC12 nuclear extract is the ubiquitously expressed transcription factor, the upstream stimulatory factor (USF). Mutation of this element by insertion of a 10 bp linker into the E box motif, in an rPPT promoter fragment spanning -865 to +92, destroys the ability of this promoter fragment to support reporter gene expression in a PC12 cell model of rPPT promoter activity. The data indicate that this rPPT E box element is likely to function as an important cis-regulatory domain in the rPPT promoter.
Paterson
J M
JM
MRC Brain Metabolism Unit, Royal Edinburgh Hospital, Scotland, U.K.
Morrison
C F
CF
Mendelson
S C
SC
McAllister
J
J
Quinn
J P
JP
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D002455:Cell Division; D002467:Cell Nucleus / Q000378:metabolism; D004268:DNA-Binding Proteins; D018274:Electroporation; D006367:HeLa Cells; D018257:Helix-Loop-Helix Motifs; D006801:Humans; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D016716:PC12 Cells; D011401:Promoter Regions, Genetic; D011498:Protein Precursors / Q000235:genetics*; D051381:Rats; D012045:Regulatory Sequences, Nucleic Acid; D015320:Tachykinins / Q000235:genetics*; D014157:Transcription Factors / Q000302:isolation & purification / Q000378:metabolism*; D051720:Upstream Stimulatory Factors
D004268:DNA-Binding Proteins; D015345:Oligonucleotide Probes; D011498:Protein Precursors; D015320:Tachykinins; D014157:Transcription Factors; C497181:USF1 protein, human; D051720:Upstream Stimulatory Factors; C497182:Usf1 protein, rat; C044206:preprotachykinin