DNA elements of the type 1 adenylyl cyclase gene locus enhance reporter gene expression in neurons and pinealocytes.
The Ca2+-stimulated type 1 adenylyl cyclase (AC1) contributes to several forms of synaptic plasticity and is the only known neurospecific adenylyl cyclase. Furthermore, the protein and mRNA levels of AC1 undergo a circadian oscillation in the pineal gland, and AC1 may play a pivotal role in regulating nocturnal melatonin synthesis. To better understand the expression of AC1, we isolated mouse genomic DNA clones of AC1. The transcription and translation start regions of mouse AC1 share extensive homologies with the bovine counterpart. The upstream proximal region has potential binding sites for transcription factors, including the steroid receptor family, the E-box factors, and Sp1. A 280-bp fragment that contains the transcription start site directed reporter gene expression in cultured cortical neurons and pinealocytes functioning as a basal neuro- and pineal-directed promoter. Interestingly, pinealocyte expression of the reporter gene was inhibited by increases in cAMP. This cAMP sensitivity may explain why AC1 mRNA in the pineal is low at night when cAMP is elevated and high during the day when cAMP signals drop. An adjacent 330-bp fragment interacted specifically with nuclear factor(s) that we designate binary E-box factor (BEF). Methylation interference and DNase I footprinting identified the BEF-binding site sequence as 5'-CCAAGGTCACGTGGC-3'. When linked to the basal tissue-directed promoter, this 15-bp sequence further enhanced reporter expression in neurons and pinealocytes. We propose that this 15-bp sequence may contribute to increased expression of AC1 in neurons and pinealocytes relative to other cells.
Chan
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Department of Pharmacology, Box 357280, University of Washington, Seattle, WA 98195, USA.
Lernmark
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D000262:Adenylyl Cyclases / Q000235:genetics*; D000818:Animals; D000831:Animals, Newborn; D001483:Base Sequence / Q000502:physiology; D001665:Binding Sites / Q000235:genetics; D001667:Binding, Competitive / Q000235:genetics; D001921:Brain / Q000166:cytology / Q000201:enzymology*; D002478:Cells, Cultured / Q000166:cytology / Q000378:metabolism; D000242:Cyclic AMP / Q000235:genetics / Q000378:metabolism; D004247:DNA / Q000737:chemistry / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology*; D017930:Genes, Reporter / Q000502:physiology*; D006150:Guanine Nucleotides / Q000378:metabolism; D008969:Molecular Sequence Data; D009474:Neurons / Q000166:cytology / Q000201:enzymology*; D009841:Oligonucleotides / Q000378:metabolism; D010870:Pineal Gland / Q000166:cytology / Q000201:enzymology*; D014176:Protein Biosynthesis / Q000502:physiology; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D020218:Response Elements / Q000502:physiology; D015398:Signal Transduction / Q000235:genetics; D016385:TATA Box / Q000502:physiology; D014157:Transcription Factors / Q000502:physiology; D014158:Transcription, Genetic / Q000502:physiology
D004268:DNA-Binding Proteins; D006150:Guanine Nucleotides; D009841:Oligonucleotides; D012333:RNA, Messenger; D014157:Transcription Factors; D004247:DNA; D000242:Cyclic AMP; D000262:Adenylyl Cyclases; C106670:adenylyl cyclase 1