Molecular cloning of complementary DNA encoding the avian receptor for vitamin D

Donald P. Mcdonnell, David J. Mangelsdorf, J. Wesley Pike, Mark R. Haussler, Bert W. O'Malley

Research output: Contribution to journalArticle

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Abstract

Vitamin D3 are intracellular proteins that mediate the nuclear action of the active metabolite 1,25-dihydroxyvitamin D3 [1,25(OH)2D3]. Two receptor-specific monoclonal antibodies were used to recover the complementary DNA (cDNA) of this regulatory protein from a chicken intestinal λgt11 cDNA expression library. The amino acid sequences that were deduced from this cDNA revealed a highly conserved cysteine-rich region that displayed homology with a domain characteristic of other steroid receptors and with gag-erbA oncogene product of avian erythroblastosis virus. RNA selected via hybridization with this DNA sequence directed the cell-free synthesis of immunopreapitable vitamin D3 receptor. Northern blot analysis of polyadenylated RNA with these cDNA probes revealed two vitamin D receptor messenger RNAs (mRNAs) of 2.6 and 3.2 kilobases in receptor-containing chicken tissues and a major cross-hybridizing receptor mRNA species of 4.2 kilobases in mouse 3T6 fibroblasts. The 4.2-kilobase species was substantially increased by prior exposure of 3T6 cells to 1,25(OH) 2D3. This cDNA represents perhaps the rarest mRNA cloned to date in eukaryotes, as well as the first receptor sequence described for an authentic vitamin.

Original languageEnglish (US)
Pages (from-to)1214-1217
Number of pages4
JournalScience
Volume235
Issue number4793
StatePublished - 1987

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Calcitriol Receptors
Molecular Cloning
Complementary DNA
Messenger RNA
Chickens
Alpharetrovirus
Calcitriol
Steroid Receptors
Cholecalciferol
Oncogene Proteins
DNA Probes
Nuclear Proteins
Eukaryota
Gene Library
Vitamins
Northern Blotting
Cysteine
Amino Acid Sequence
Fibroblasts
Monoclonal Antibodies

ASJC Scopus subject areas

  • General

Cite this

Mcdonnell, D. P., Mangelsdorf, D. J., Pike, J. W., Haussler, M. R., & O'Malley, B. W. (1987). Molecular cloning of complementary DNA encoding the avian receptor for vitamin D. Science, 235(4793), 1214-1217.

Molecular cloning of complementary DNA encoding the avian receptor for vitamin D. / Mcdonnell, Donald P.; Mangelsdorf, David J.; Pike, J. Wesley; Haussler, Mark R.; O'Malley, Bert W.

In: Science, Vol. 235, No. 4793, 1987, p. 1214-1217.

Research output: Contribution to journalArticle

Mcdonnell, DP, Mangelsdorf, DJ, Pike, JW, Haussler, MR & O'Malley, BW 1987, 'Molecular cloning of complementary DNA encoding the avian receptor for vitamin D', Science, vol. 235, no. 4793, pp. 1214-1217.
Mcdonnell DP, Mangelsdorf DJ, Pike JW, Haussler MR, O'Malley BW. Molecular cloning of complementary DNA encoding the avian receptor for vitamin D. Science. 1987;235(4793):1214-1217.
Mcdonnell, Donald P. ; Mangelsdorf, David J. ; Pike, J. Wesley ; Haussler, Mark R. ; O'Malley, Bert W. / Molecular cloning of complementary DNA encoding the avian receptor for vitamin D. In: Science. 1987 ; Vol. 235, No. 4793. pp. 1214-1217.
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N2 - Vitamin D3 are intracellular proteins that mediate the nuclear action of the active metabolite 1,25-dihydroxyvitamin D3 [1,25(OH)2D3]. Two receptor-specific monoclonal antibodies were used to recover the complementary DNA (cDNA) of this regulatory protein from a chicken intestinal λgt11 cDNA expression library. The amino acid sequences that were deduced from this cDNA revealed a highly conserved cysteine-rich region that displayed homology with a domain characteristic of other steroid receptors and with gag-erbA oncogene product of avian erythroblastosis virus. RNA selected via hybridization with this DNA sequence directed the cell-free synthesis of immunopreapitable vitamin D3 receptor. Northern blot analysis of polyadenylated RNA with these cDNA probes revealed two vitamin D receptor messenger RNAs (mRNAs) of 2.6 and 3.2 kilobases in receptor-containing chicken tissues and a major cross-hybridizing receptor mRNA species of 4.2 kilobases in mouse 3T6 fibroblasts. The 4.2-kilobase species was substantially increased by prior exposure of 3T6 cells to 1,25(OH) 2D3. This cDNA represents perhaps the rarest mRNA cloned to date in eukaryotes, as well as the first receptor sequence described for an authentic vitamin.

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