Puralpha, a single-stranded DNA binding protein, suppresses the enhancer activity of cAMP response element (CRE).
Puralpha, a single-stranded DNA binding protein, recognizes a PUR element (GGN repeat). We have reported that Puralpha binds to a single-stranded oligonucleotide probe containing the cAMP response element (CRE) of rat somatostatin gene using a gel mobility shift assay. Here, we showed that Puralpha binds to the probe only in the presence of a PUR element by a more detailed characterization. We also examined the effects of Puralpha on the enhancer activity of the somatostatin CRE in PC12 cells using the reporter gene assay. Transfected Puralpha suppressed the CRE enhancer activity stimulated by forskolin (which increases intracellular cAMP), but suppression was not observed when the PUR element was deleted. The neurite extension induced by forskolin was inhibited by the transfection of Puralpha, but that by NGF was not suppressed. The c-fos mRNA induced by forskolin, but not by NGF, was also suppressed by Puralpha transfection. These results indicate that Puralpha suppresses the biological activities induced by forskolin, but not by NGF, in PC12 cells and that Puralpha could interfere with a cAMP-CRE signal pathway.
Sadakata
T
T
Department of Pharmacology A6, Osaka University Medical School, 2-2 Yamadaoka, Suita, Japan.
Kuo
C
C
Ichikawa
H
H
Nishikawa
E
E
Niu
S Y
SY
Kumamaru
E
E
Miki
N
N
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D005576:Colforsin / Q000494:pharmacology; D017362:Cyclic AMP Response Element-Binding Protein / Q000378:metabolism*; D004268:DNA-Binding Proteins; D004742:Enhancer Elements, Genetic / Q000187:drug effects; D017930:Genes, Reporter; D007700:Kinetics; D008156:Luciferases / Q000235:genetics; D015345:Oligonucleotide Probes; D016716:PC12 Cells; D051381:Rats; D013004:Somatostatin / Q000235:genetics*; D013489:Suppression, Genetic; D014157:Transcription Factors; D014162:Transfection; D018911:Trinucleotide Repeats
D017362:Cyclic AMP Response Element-Binding Protein; D004268:DNA-Binding Proteins; D015345:Oligonucleotide Probes; D014157:Transcription Factors; D005576:Colforsin; D013004:Somatostatin; D008156:Luciferases