Characterization of the regulatory region of Adra2c, the gene encoding the murine alpha2C adrenoceptor subtype.
The 5' flanking sequence (3,227 base pairs, bp) of the mouse Adra2c subtype gene was determined and characterized. The transcription start site was mapped to nucleotide 'A' of two initiator motifs in tandem array, i.e. 1,159 and 1,153 bp upstream from the initiation codon of the open reading frame (ORF) of Adra2c, respectively. One structural feature salient to the 5' regulatory region of Adra2c is present in the sequence 1 kb immediately upstream from the receptor ORF, which is highly enriched in GC content (76%) and CpG island counts (i.e. CpG/GpC, 146:177), and thus rich in Sp1-binding motifs. At the 3' flanking region, the polyadenylation signal was mapped to 481 bp downstream from the termination codon. The transcript defined by sequence data thereby is consistent with a size of 3 kb (brain form) determined by Northern blot analysis. The transgene, Adra2c-NN- lacZ, which links the promoter region of Adra2c to the lacZ reporter gene, was constructed in order to evaluate the functional capacity of the promoter and the putative motifs residing within the defined regulatory region (1.9 kb upstream from the ORF) in directing the reporter gene expression in vitro in transiently transfected cells and in vivo in transgenic (Tg) mice. Permissive cell types to Adra2c-NN include those derived from neural and kidney lineages. Significant Adra2c-NN-driven reporter expression in Tg mice established suggests that alpha2C adrenoceptor expression is permissive under Adra2c-NN in central (cerebral cortex, hippocampus, subthalamus, hypothalamus, superior colliculus, cerebellum, and brain stem) and peripheral (pancreatic beta-islets) tissues.
Chen
Wen-Kwei
WK
Institute of Microbiology and Immunology, National Yang-Ming University, Taipei, ROC.
Chang
Nan-Chi A
NC
Chang
Yen-Hwa
YH
Chang
Kuo-Long
KL
Wu
Shinn-Chih
SC
Yang
Tzong-Shang
TS
Wu
Sheue-Mei
SM
Chang
Alice Chien
AC
D020816:Amino Acid Motifs; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D001921:Brain / Q000378:metabolism; D002490:Central Nervous System / Q000378:metabolism; D003001:Cloning, Molecular; D003062:Codon; D019295:Computational Biology; D018899:CpG Islands; D015723:Gene Library; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D006801:Humans; D007515:Islets of Langerhans / Q000378:metabolism; D007763:Lac Operon; D051379:Mice; D008822:Mice, Transgenic; D008957:Models, Genetic; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D016366:Open Reading Frames; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D018341:Receptors, Adrenergic, alpha-2 / Q000737:chemistry* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014018:Tissue Distribution; D014157:Transcription Factors; D014158:Transcription, Genetic; D014162:Transfection; D019076:Transgenes; D001616:beta-Galactosidase / Q000378:metabolism
C547774:ADRA2C protein, human; C547775:Adra2c protein, mouse; D003062:Codon; D018341:Receptors, Adrenergic, alpha-2; D014157:Transcription Factors; D001616:beta-Galactosidase