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PMID: 8668154 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Intron retention generates a novel isoform of the murine vitamin D receptor that acts in a dominant negative way on the vitamin D signaling pathway.

Molecular and cellular biology ·Vol. 16 ·No. 7 ·1996-07-00 ·Pages 3393-400

Ebihara K, Masuhiro Y, Kitamoto T, Suzawa M, Uematsu Y, Yoshizawa T, Ono T, Harada H, Matsuda K, Hasegawa T, Masushige S, Kato S

Abstract

We identified and characterized a novel rat vitamin D receptor isoform (rVDR1), which retains intron 8 of the canonical VDR (rVDR0) during alternative splicing. In this isoform protein directed by the stop codon in this newly identified exon, a part of the ligand binding domain (86 amino acids) is truncated at the C-terminal end but contains 19 extra amino acids. The rVDR1 transcript was expressed at a level 1/15 to 1/20 of that of rVDR0 in the kidney and intestine in adult rats but not in embryos. The recombinant rVDR1 protein showed no ligand binding activity. Homo- and heterodimers of the recombinant rVDR0 and rVDR1 proteins bound to a consensus vitamin D response element (VDRE) but not to consensus response elements for thyroid hormone and retinoic acid. However, unlike rVDR0, rVDR1 did not form a heterodimeric complex with RXR on the VDRE. A transient expression assay showed that this isoform acted as a dominant negative receptor against rVDR0 transactivation. Interestingly, the dominant negative activities of rVDR1 differed among VDREs. Thus, the present study indicates that this new VDR isoform negatively modulates the vitamin D signaling pathway, through a particular set of target genes.

MeSH Terms
Alternative Splicing Amino Acid Sequence Animals Base Sequence Calcitriol/metabolism Chloramphenicol O-Acetyltransferase/biosynthesis Embryo, Mammalian HeLa Cells Humans Intestinal Mucosa/metabolism Introns Kidney/metabolism Molecular Sequence Data Oligonucleotide Probes Polymerase Chain Reaction Rats Receptors, Calcitriol/biosynthesis,genetics,physiology Receptors, Retinoic Acid/physiology Recombinant Fusion Proteins/metabolism Restriction Mapping Retinoid X Receptors Signal Transduction Transcription Factors/physiology Transcription, Genetic Transfection Vitamin D/metabolism
Chemicals
Oligonucleotide Probes Receptors, Calcitriol Receptors, Retinoic Acid Recombinant Fusion Proteins Retinoid X Receptors Transcription Factors Vitamin D Chloramphenicol O-Acetyltransferase Calcitriol
Authors & Affiliations
12 authors, click to expand affiliations / ORCID
Ebihara K
Department of Agricultural Chemistry, Tokyo University of Agriculture, Setagayaku, Japan.
Masuhiro Y
Kitamoto T
Suzawa M
Uematsu Y
Yoshizawa T
Ono T
Harada H
Matsuda K
Hasegawa T
Masushige S
Kato S
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.16.7.3393
Published
1996-07-00
Pages
3393-400
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC231333
Subset
IM
External Links
PubMed source DOI: 10.1128/MCB.16.7.3393 PMC Full Text View journal details
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