Protein/DNA interaction profiling reveals novel regulators of the pineal transcriptome.
The rat pineal gland transcriptome exhibits dynamic daily variation that reflects nocturnally restricted hormone production. Here we have used a protein/DNA interaction array to screen for day-night changes in DNA binding activity that are associated with transcriptional rhythms. Overall, 47 of 54 potential consensus binding sequence activities were detected, and of these, 29 (62%) were found to exhibit day:night differences in level. In addition to known, rhythmic pineal DNA binding activities (CRE and AP-1), multiple novel activities were observed including nocturnally elevated AP-2 consensus sequence binding activity. This array result was validated using conventional DNA binding assays, and we have also demonstrated AP-2beta and AP-2gamma proteins in the pineal gland, in addition to a nocturnally elevated AP-2alpha isoform. Our results have confirmed the presence of a complex assembly of transcriptional rhythms in the rat pineal gland and have provided details of more factors that contribute to this aspect of circadian neuroendocrine function.
Davies
J S
JS
School of Biosciences, Cardiff University, Cardiff CF10 3US, UK.
Carter
D A
DA
D000818:Animals; D001665:Binding Sites; D004247:DNA / Q000235:genetics* / Q000378:metabolism; D017931:DNA Primers; D049449:Luminescence; D020411:Oligonucleotide Array Sequence Analysis; D010870:Pineal Gland / Q000502:physiology*; D011506:Proteins / Q000235:genetics*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012160:Retina / Q000502:physiology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014158:Transcription, Genetic
D017931:DNA Primers; D011506:Proteins; D004247:DNA