Linked Life Data

12039974

Source:http://linkedlifedata.com/resource/pubmed/id/12039974

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
6
pubmed:dateCreated
2002-5-31
pubmed:abstractText
The active form of vitamin D is synthesized by 25-hydroxyvitamin D 1alpha-hydroxylase (1alpha-hydroxylase), which is expressed predominantly in renal proximal tubular cells. To clarify the mechanism of cell-specific gene expression of this enzyme, the 5'-flanking region of the mouse 1alpha-hydroxylase gene was investigated. Investigation began with mRNA expression of 1alpha-hydroxylase in cultured cells, including LLC-PK1, NIH/3T3, HepG2, MDCK, and OK cells. Expression of 1alpha-hydroxylase mRNA was restricted in LLC-PK1 cells. Several lengths of the 5'-flanking region of 1alpha-hydroxylase gene were linked to a pGL3-basic luciferase vector and introduced into these cells. Only LLC-PK1 cells had a substantial luciferase activity. Deletion analyses revealed that luciferase activity was detected in constructs extending from the transcription initiation site to -1652 to -105 bp, whereas further deletion to -80 bp resulted in a marked decrease in activity. The region from -105 to -80 bp contained two ternary complex factor-1 (TCF-1) sites, and mutations in the proximal TCF-1 site decreased the activity. Electrophoretic mobility shift assay demonstrated binding of LLC-PK1 nuclear proteins to this region. Tests of enhancer function in LLC-PK1 cells indicated that the 26-bp fragment behaved as a classical enhancer, i.e., independently of position and orientation. Moreover, a decoy oligonucleotide corresponding to this region substantially inhibited the promoter activity of 1alpha-hydroxylase gene. This study suggests that the -105 to -80 bp element of mouse 1alpha-hydroxylase gene contains an enhancer to be necessary for renal proximal tubular cell-specific expression.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Jun
pubmed:issn
1046-6673
pubmed:author
pubmed:issnType
Print
pubmed:volume
13
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
1455-63
pubmed:dateRevised
2008-11-21
pubmed:meshHeading
pubmed:year
2002
pubmed:articleTitle
Identification of a renal proximal tubular cell-specific enhancer in the mouse 25-hydroxyvitamin d 1alpha-hydroxylase gene.
pubmed:affiliation
Department of Internal Medicine, School of Medicine, Keio University, Tokyo, Japan.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't