Brain-specific promoter/exon I.f of the cyp19a1 (aromatase) gene in Xenopus laevis.
Aromatase, encoded by the cyp19a1 gene, is the key enzyme for estrogen biosynthesis. Exon I.f of aromatase transcripts in the Xenopus brain is driven in a brain-specific manner. In this study, we cloned brain aromatase with a 5'-end of various lengths by 5'-RACE and detected the expression pattern of the aromatase mRNA. In Xenopus at the larval stage, the brain aromatase mRNA expression was five-fold higher than those in the gonad and liver, and was upregulated from stage 42 to stage 50. After isolating the brain-specific promoter I.f, which was located ∼6.5 kb upstream from gonad-specific exon PII, we observed this promoter in a potential cis-elements for several transcriptional factors, such as Oct-1, c-Myc, the GATA gene family, C/EBPalpha, Sox5, p300, XFD-1, AP1, the STAT gene family, FOXD3, and the Smad gene family. In addition, the core promoter elements of two initiators and an atypical TATA box were found around the 5'-RACE products. In the 5'-flanking region of exon I.f, the binding sites for nuclear extracts suggested that the followings are important: the STAT gene family, a 38-bp conserved region among five species, FOXD3, and the Smad gene family within the region 200 bp upstream from the transcription initiation site. Real-time RT-PCR analysis showed that the foxd3, smad2 and smad4.1/4.2 mRNAs are specifically expressed in the brain. Furthermore, the expression change of foxd3, which has been reported as a repressor, indicated that expression decreased to stage 50 from stage 42, contrary to that of aromatase mRNA. These results may imply that foxd3 expression decreases and aromatase expression increases as a result of the contribution to promoter I.f by transcriptional activators such as smads. However, since these putative cis-elements and transcription initiation sites are not conserved in the brain-specific promoter of other species, this transcriptional regulatory mechanism of exon I.f may be characteristic of Xenopus.
Laboratory of Biochemistry, Department of Chemistry, College of Humanities and Sciences, Nihon University, 3-25-40 Sakurajosui, Setagaya-ku, Tokyo 156-8550, Japan.
Tadahiko
T
Nakagawa
Junshin
J
Iwabuchi
D000818:Animals; D001141:Aromatase / Q000235:genetics* / Q000378:metabolism; D001921:Brain / Q000502:physiology*; D003850:Deoxyribonuclease I / Q000235:genetics / Q000378:metabolism; D005091:Exons; D051858:Forkhead Transcription Factors / Q000235:genetics / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic; D007438:Introns; D009928:Organ Specificity; D011401:Promoter Regions, Genetic; D012045:Regulatory Sequences, Nucleic Acid; D016385:TATA Box; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D024363:Transcription Initiation Site; D029867:Xenopus Proteins / Q000235:genetics* / Q000378:metabolism; D014982:Xenopus laevis / Q000235:genetics
D051858:Forkhead Transcription Factors; D014157:Transcription Factors; D029867:Xenopus Proteins; C491003:foxd3b-A protein, Xenopus; D001141:Aromatase; D003850:Deoxyribonuclease I