Structural features of the murine gene encoding the RI beta subunit of cAMP-dependent protein kinase.
The activation of cyclic AMP-dependent protein kinase is controlled by the regulatory (R) subunits of the holoenzyme. Here we present a characterization of the mouse RI beta subunit gene, which in contrast to other subunit genes of cyclic AMP-dependent protein kinase is expressed almost exclusively in neurons. It was determined that RI beta is relatively large with 11 exons spanning a minimum 75 kb. The mouse chromosomal locus (designated Prkar1b) was determined by interspecific backcross mapping and found to reside on the distal arm of chromosome 5. Previously, it was shown that 3.5 kb of DNA encompassing the RI beta promoter could direct neural-specific gene expression in transgenic mice. Analysis of this DNA suggests the presence of an unusually large number of binding sites for transcription factors ranging from tissue-specific regulators, immediate-early genes, and mediators of hormone action. In addition to 18 putative SP1 sites, we identified 27 consensus sequences for basic Helix-Loop-Helix, POU, and Pax family members, 5 AP1 sites, and over 40 half-sites for the superfamily of steroid hormone receptor. Gel mobility-shift assays employing brain nuclear extract and pure transcription factor protein established that many of these DNA sequences are functional in binding protein. The abundance and configuration of transcription factor binding sites within the promoter region of RI beta suggests that this gene is subject to complex modes of regulation in neurons.
Bristol-Myers Squibb Pharmaceutical Research Institute, Seattle, Washington 98121.
C H
CH
Clegg
N R
NR
Koeiman
N A
NA
Jenkins
D J
DJ
Gilbert
N G
NG
Copeland
M G
MG
Neubauer
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D002874:Chromosome Mapping; D017868:Cyclic AMP-Dependent Protein Kinases / Q000235:genetics* / Q000378:metabolism; D004247:DNA / Q000235:genetics; D018257:Helix-Loop-Helix Motifs; D006728:Hormones / Q000378:metabolism; D051379:Mice / Q000235:genetics*; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes / Q000235:genetics; D011401:Promoter Regions, Genetic; D016755:Proto-Oncogene Proteins c-jun / Q000378:metabolism; D011987:Receptors, Steroid / Q000378:metabolism; D012091:Repetitive Sequences, Nucleic Acid; D014157:Transcription Factors / Q000378:metabolism
D006728:Hormones; D015345:Oligonucleotide Probes; D016755:Proto-Oncogene Proteins c-jun; D011987:Receptors, Steroid; D014157:Transcription Factors; D004247:DNA; D017868:Cyclic AMP-Dependent Protein Kinases