Chromatin structure of the bovine Cyp19 promoter 1.1. DNaseI hypersensitive sites and DNA hypomethylation correlate with placental expression.
Expression of the Cyp19 gene, encoding aromatase cytochrome P450, is driven by several tissue-specific promoters. The underlying mechanisms of this complex regulation have not yet been elucidated in detail. In the present report we investigate a possible link between chromatin structure and tissue-specific regulation of the bovine Cyp19 gene. We analysed the DNA methylation status and mapped DNaseI hypersensitive sites in the region encompassing the Cyp19 promoter 1.1 (P1.1) which controls Cyp19 expression in the bovine placenta. We show that P1.1 is hypomethylated in placental cotyledons (foetal layer) whereas it is methylated in placental caruncles (maternal layer), testis and corpus luteum. Furthermore, two placenta-specific DNaseI hypersensitive sites, HS1 and HS2, were observed within P1.1. Both DNA hypomethylation and the presence of DNaseI hypersensitive sites correlate with transcriptional activity of P1.1. Sequence analysis of hypersensitive sites revealed potential cis-regulatory elements, an E-box in HS1 and a trophoblast-specific element-like sequence in HS2. It could be demonstrated by electrophoretic mobility shift assays that both sequence motifs are specific targets for placenta-derived nuclear factors. In conclusion, observed tissue-specific differences of the chromatin structure which correlate with tissue-specific promoter activity suggest that chromatin might be an important regulator of aromatase expression in cattle.
Fürbass
R
R
Research Unit Molecular Biology, Research Institute for the Biology of Farm Animals, Dummerstorf, Germany. fuerbas@fbn-dummerstorf.de
Said
H M
HM
Schwerin
M
M
Vanselow
J
J
D000818:Animals; D001141:Aromatase / Q000235:genetics*; D001665:Binding Sites; D002417:Cattle; D002843:Chromatin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D019175:DNA Methylation; D015342:DNA Probes / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism; D003850:Deoxyribonuclease I / Q000378:metabolism*; D005786:Gene Expression Regulation; D009687:Nuclear Proteins / Q000378:metabolism; D023282:Nuclease Protection Assays; D009928:Organ Specificity; D010920:Placenta / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D020218:Response Elements / Q000235:genetics; D014158:Transcription, Genetic
D002843:Chromatin; D015342:DNA Probes; D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; D001141:Aromatase; D003850:Deoxyribonuclease I