DNA sequences in the rat parathyroid hormone-related peptide gene responsible for 1,25-dihydroxyvitamin D-3-mediated transcriptional repression

被引:64
作者
Falzon, M [1 ]
机构
[1] UNIV TEXAS, MED BRANCH, SEALY CTR MOL SCI, GALVESTON, TX 77555 USA
关键词
D O I
10.1210/me.10.6.672
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
Expression of the gene encoding PTH-related peptide (PTHrP), a protein that plays a primary role in the development of humoral hypercalcemia of malignancy, is down-regulated at the transcriptional level by 1,25-dihydroxyvitamin D-3 [1,25-(OH)(2)D-3]. Deletions of the 5'-flanking region of the rat PTHrP gene, when fused to the chloramphenicol acetyltransferase gene and transfected into ROS 17/2.8 (rat osteosarcoma) cells, showed that the 1,25-(OH)(2)D-3 responsive region is located between -1.05 and -0.71 kb upstream of the transcription start site. Further mapping of this region revealed that a 123-bp fragment is able to confer 1,25-(OH)(2)D-3 responsiveness to a heterologous (SV40) promoter. This region contains two potential vitamin D response elements (VDREs). One of these motifs resembles the negative VDRE (nVDRE) from the PTH gene, which is also down-regulated by vitamin D-3. The other element resembles the canonical VDRE (two hexanucleotide motifs separated by three nucleotides), which has been characterized in a number of genes whose expression is modulated by vitamin D-3. Electrophoretic mobility shift assays using nuclear extracts from ROS 17/2.8 cells and from vitamin D receptor (VDR)-enriched COS 1 cells revealed that both elements interact with the VDR. This protein-DNA interaction is disrupted by an anti-VDR antibody. Therefore, modulation of PTHrP gene transcription by 1,25-(OH)(2)D-3 is mediated by the VDR interacting with one or both of the identified motifs in the 5'-flanking sequence of the gene.
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页码:672 / 681
页数:10
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