Characterization of the 5′ flanking region of the rat D3 dopamine receptor gene

Ursula M. D'Souza, Wei Wang, Da Qing Gao, Seiji Kanda, Gwang Lee, Eunsung Junn, Cheol Kyu Hwang, Pedro A. Jose, M. Maral Mouradian

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17 Scopus citations


The D3 dopamine receptor has a restricted regional distribution in brain and is regulated by dopaminergic agents. Additionally, the D3 gene is implicated in the pathogenesis of several neuropsychiatric disorders or in their response to pharmacological agents. Elucidating its transcription control mechanisms is therefore of interest in order to explain these biological features of the D3 gene. In this study, the 5′ flanking region of the rat D3 gene was characterized by isolating the 5′ end of its cDNA as well as 4.6 kb of genomic sequence. Analysis of this region revealed the presence of two new exons 196-bp and 120-bp long, separated by an 855-bp intron, located several kilobases upstream of the previously published coding exons. Thus, current evidence indicates that the rat D3 gene is organized into eight exons. Transcription initiation site was determined by primer extension analysis and repeated rounds of 5′ RACE and was found to localize at a pyrimidine-rich consensus 'initiator' sequence, similar to the rat D2 gene. The D3 promoter lacks TATA or CAAT boxes but unlike that of other dopamine receptor genes has only 52% GC content. Functional analysis of D3 promoter deletion mutants fused to a reporter gene in TE671 cells, which endogenously express this gene, revealed strong transcriptional activity localized within 36 nucleotides upstream of transcription start site, and a potent silencer between bases -37 and -537. The D3 promoter is inactive in C6 and COS7 cells. We conclude that the D3 gene, similar to the closely related D2 gene, is transcribed from a tissue specific promoter which is under intense negative control.

Original languageEnglish (US)
Pages (from-to)1736-1744
Number of pages9
JournalJournal of Neurochemistry
Issue number6
StatePublished - 2001


  • 5′ Flanking region
  • D gene
  • Dopamine receptor gene
  • Promoter region
  • Transcription regulation


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